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Chapter 12 principles of diagnosis and prevention of pathogen infection. Bacteriological diagnosis※ Virology diagnosis※ Mycology diagnosis Specific prophylaxis and therapy. section 1 bacterial diagnosis bacteriological diagnosis: - PowerPoint PPT Presentation
Citation preview
Chapter 12Chapter 12
principles of principles of diagnosis and diagnosis and prevention of prevention of pathogen infectionpathogen infection
Bacteriological diagnosisBacteriological diagnosisVirology diagnosisVirology diagnosisMycology diagnosisMycology diagnosisSpecific prophylaxis and therapySpecific prophylaxis and therapy
section 1 bacterial diagnosissection 1 bacterial diagnosis
bacteriological diagnosisbacteriological diagnosischeck pathogen and their componentscheck pathogen and their components
(( Ag products nucleic acidAg products nucleic acid )) detect Ag with known Abmdash detect Ag with known Abmdash serological identificationserological identificationdetect Ab in serum detect Ab in serum detect unknown Ab with known Agmdashdetect unknown Ab with known Agmdash serodiagnosisserodiagnosis
Steps precede the laboratory Steps precede the laboratory workwork
11 Choose the appropriate specimenChoose the appropriate specimen
22 Obtain the specimen avoid contaminatedObtain the specimen avoid contaminated
33 Transport the specimen promptly or Transport the specimen promptly or store it correctlystore it correctly
44 Provide essential informationProvide essential information
(二)(二) detection of pathogenic bacteria detection of pathogenic bacteria
11 morphology and structuremorphology and structure
22 isolating cultureisolating culture
33 detection of Agdetection of Ag
44 other methodsother methods
11 morphosis detectionmorphosis detection
procedureprocedure smear fix stain observe under smear fix stain observe under
microscopemicroscope
(( shapesizearrangement staining shapesizearrangement staining special structurespecial structure ) ) identify bacteriaidentify bacteria
major technique for detection of pathogmajor technique for detection of pathogenic bacteriaenic bacteria
(( 11 )) microscopemicroscope
Alight microscopeAlight microscope
resolutionresolution 025μm025μm
magnificationmagnification times1000times1000
B electronmicroscopeB electronmicroscope
resolutionresolution 1nm
magnification times100000
--cannot observe viable organism--cannot observe viable organism
C Other microscopeC Other microscope
bullbulldarkfield microscopedarkfield microscope
bull bullcontrast phase microscopecontrast phase microscope
bull bullfluorescence microscopefluorescence microscope
bull bullconfocal microscopeconfocal microscope
(( 22 )) staining techniquestaining technique
simple stainingsimple staining only use one dye only use one dye observe morphology arrangemen observe morphology arrangementt
compound stainingcompound staining use 2 kinds dyes or use 2 kinds dyes or more to identify bacteria more to identify bacteria
Gram stainingGram staining positivepositive (( violetviolet )) negativenegative (( redred ))Acid-fast stainingAcid-fast staining positivepositive (( redred )) negative negative (( blueblue ))Special stainingSpecial staining for bacterial capsule for bacterial capsule
spore flagella and so onspore flagella and so on
22 isolating culture and identifyisolating culture and identifyGeneral procedureGeneral procedure
specimen
Plain agar
Blood agar
Selective medium
cultural feature
nutrition
growing condition
Colony feature
hemolysis
pure
inoculate
stain initial identify (morphology)
Biochemical reaction
Serological identificationSerological identification species groupspecies groupss
Virulence Virulence identificationidentification
detect endotoxin or exotoxindetect endotoxin or exotoxin
animal testanimal test rabbit guinea pirabbit guinea pigg
Susceptibility Susceptibility testtest
Slip methodSlip method determine the naturedetermine the nature
Test tube methodTest tube method quantitationquantitation(( detect MBCdetect MBC MICMIC ))
33 detect Agdetect Ag
mdashmdash mdashmdashserological identificationserological identification
highly specificityhighly specificity
sensitivity sensitivity
44 other detectionother detection
Detect metabolic productDetect metabolic product
bull bull PCR detect DNAPCR detect DNA
caution avoid contaminationcaution avoid contamination (faulse positive)(faulse positive)
bull bull DNA chips techniqueDNA chips technique
22 serological diagnosisserological diagnosis
-----detect Ab with known Ag-----detect Ab with known Ag
------specimens------specimens serumserum
------paired serum (acute------paired serum (acute convalescent serum)convalescent serum)
Compared acute serum and convalescenCompared acute serum and convalescent serum quantitativelyt serum quantitatively
A 4-fold or greater increase in Ab titerA 4-fold or greater increase in Ab titer su supports a diagnosis of recent infectionpports a diagnosis of recent infection
Serum containing a high titer of Abs of thSerum containing a high titer of Abs of thee IgM IgM subclass would suggest a subclass would suggest a current icurrent infectionnfection
commonly used methodscommonly used methods
Agglutination testAgglutination test Widal test---diagnose typhus and Widal test---diagnose typhus and
paratyphoidparatyphoid
Well-Felix---diagnose rickettsiosisWell-Felix---diagnose rickettsiosis
10486801048680
Neutralization testNeutralization test antildquoOrdquo test---diagnose rheumatic antildquoOrdquo test---diagnose rheumatic
diseasedisease
Section 2 virology diagnosisSection 2 virology diagnosis
ViralViral infeinfectionction
Electron microscopeElectron microscope
Light microscope inLight microscope inclusion bodyclusion body
isolationisolation
VirusVirus particleparticle
V AgV Ag
V nucleic acidV nucleic acid
Viral enzymeViral enzyme retroviridaseretroviridase
DetectDetect virusvirus
DetectDetect AbAb
Choose the appropriate specimenChoose the appropriate specimen
Aseptic operationAseptic operation add antibiotics to speciadd antibiotics to speci
menmen
Take it in acute phase Take it in acute phase
Low temperature conservationLow temperature conservation transport it transport it
promptlypromptly preserve in 50preserve in 50 glycerineglycerine
Serologial diagnose paired serumSerologial diagnose paired serum
Specimen Selection Collection and ProcessingSpecimen Selection Collection and Processing
二二 viral isolation and identifyingviral isolation and identifying
specimenspecimenAnimal inoculaAnimal inoculationtion
Embryonated eEmbryonated egggg
Cell cultureCell culture
positivepositive
negativenegative
identifyingidentifying
BlindBlind passagepassage22 - 3generatiogenerationn
negativenegative
Virus negatiVirus negativeve
asepsisasepsis
(一)(一) viral isolationviral isolation
11 animal inoculationanimal inoculation
chimpanzeemonkeyrabbitratchimpanzeemonkeyrabbitrat
22 embryonated egg inoculationembryonated egg inoculation
3 Cell culture3 Cell culture (( commonly usedcommonly used ))Primary and secondary cell culturePrimary and secondary cell culture
general proceduregeneral procedure
tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell
cell monolayercell monolayer
brokenbroken proteaseprotease
(primary cell)(primary cell)Serial subcultivationSerial subcultivation
Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)
features of virus cell culturefeatures of virus cell culture
A sensitive to many kinds of virusesA sensitive to many kinds of viruses
B cost highB cost high
C carry latent virusC carry latent virus
(2)Diploid(2)Diploid cellcell cultureculture
usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine
(3) secondary cell culture(3) secondary cell culture
sensitive to sensitive to multiplemultiple viruses viruses
high reproductive capacity long genera high reproductive capacity long generation timetion time
have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine
(( 二二 )) indexindex for reproduction of virusfor reproduction of virus
(1)Virus1)Virus infected cellinfected cell
(( cell roundinggathercell roundinggather
necrosisnecrosis ))
1 Cytopathic effect CPE)1 Cytopathic effect CPE)
CPECPE
(( 22 )) cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation
(( 33 )) viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell
CPECPE
22 hemadsorption hemadsorption HAd HAd
hemagglutininhemagglutinin (( HAHA )) on membrane of vion membrane of virus infected cell rus infected cell
Can Can adsorbadsorb vertebrate vertebrate RBCRBC
3 Interference3 Interference
4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange
( 三 ) viral quantitative assayHemagglutination test total amount
Plaque bacteriophage determination
ID50 or TCID50
Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le
sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex
cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a
nd hyphand hypha
22 cultureculture
specimen sabouraud mediumspecimen sabouraud medium
macroculture microculturemacroculture microculture
colonycolony
biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique
sporespore
3animal test3animal test
4skin test hypersensitivy4skin test hypersensitivy
(三)(三) mycology rapid diagnosismycology rapid diagnosis
serological test detect Ag or Abserological test detect Ag or Ab
detect nucleic acid of fungusdetect nucleic acid of fungus
detect mycotoxindetect mycotoxin
questionsquestions
11 definitiondefinition
serological identificationserological identification
and diagnosis)and diagnosis)
Cytopathic effect CPECytopathic effect CPE
hemadsorption HAdhemadsorption HAd
interferenceinterference
22 principle of Widalrsquos testprinciple of Widalrsquos test
33 principle of Gramrsquos stainingprinciple of Gramrsquos staining
Section 4Section 4
artificial active artificial active immunityimmunity
Artificial active immunityArtificial active immunity
stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid
vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA
A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow
th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni
citycity are repeatedly selected are repeatedly selected BCGBCG
The mutation is expressed only The mutation is expressed only
under certain conditionsunder certain conditions
may be may be useful in vaccinesuseful in vaccines
Conditional-lethal mutationsConditional-lethal mutations
temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation
the organisms can the organisms can replicate replicate at a at a
relativelyrelatively low low permissive temperature permissive temperature
but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive
temperaturetemperature
Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize
d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost
errdquo injectionerrdquo injection
A Live (attenuated) A Live (attenuated) vaccinesvaccines
B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate
responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe
toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves
artificial passive artificial passive immunization immunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
Bacteriological diagnosisBacteriological diagnosisVirology diagnosisVirology diagnosisMycology diagnosisMycology diagnosisSpecific prophylaxis and therapySpecific prophylaxis and therapy
section 1 bacterial diagnosissection 1 bacterial diagnosis
bacteriological diagnosisbacteriological diagnosischeck pathogen and their componentscheck pathogen and their components
(( Ag products nucleic acidAg products nucleic acid )) detect Ag with known Abmdash detect Ag with known Abmdash serological identificationserological identificationdetect Ab in serum detect Ab in serum detect unknown Ab with known Agmdashdetect unknown Ab with known Agmdash serodiagnosisserodiagnosis
Steps precede the laboratory Steps precede the laboratory workwork
11 Choose the appropriate specimenChoose the appropriate specimen
22 Obtain the specimen avoid contaminatedObtain the specimen avoid contaminated
33 Transport the specimen promptly or Transport the specimen promptly or store it correctlystore it correctly
44 Provide essential informationProvide essential information
(二)(二) detection of pathogenic bacteria detection of pathogenic bacteria
11 morphology and structuremorphology and structure
22 isolating cultureisolating culture
33 detection of Agdetection of Ag
44 other methodsother methods
11 morphosis detectionmorphosis detection
procedureprocedure smear fix stain observe under smear fix stain observe under
microscopemicroscope
(( shapesizearrangement staining shapesizearrangement staining special structurespecial structure ) ) identify bacteriaidentify bacteria
major technique for detection of pathogmajor technique for detection of pathogenic bacteriaenic bacteria
(( 11 )) microscopemicroscope
Alight microscopeAlight microscope
resolutionresolution 025μm025μm
magnificationmagnification times1000times1000
B electronmicroscopeB electronmicroscope
resolutionresolution 1nm
magnification times100000
--cannot observe viable organism--cannot observe viable organism
C Other microscopeC Other microscope
bullbulldarkfield microscopedarkfield microscope
bull bullcontrast phase microscopecontrast phase microscope
bull bullfluorescence microscopefluorescence microscope
bull bullconfocal microscopeconfocal microscope
(( 22 )) staining techniquestaining technique
simple stainingsimple staining only use one dye only use one dye observe morphology arrangemen observe morphology arrangementt
compound stainingcompound staining use 2 kinds dyes or use 2 kinds dyes or more to identify bacteria more to identify bacteria
Gram stainingGram staining positivepositive (( violetviolet )) negativenegative (( redred ))Acid-fast stainingAcid-fast staining positivepositive (( redred )) negative negative (( blueblue ))Special stainingSpecial staining for bacterial capsule for bacterial capsule
spore flagella and so onspore flagella and so on
22 isolating culture and identifyisolating culture and identifyGeneral procedureGeneral procedure
specimen
Plain agar
Blood agar
Selective medium
cultural feature
nutrition
growing condition
Colony feature
hemolysis
pure
inoculate
stain initial identify (morphology)
Biochemical reaction
Serological identificationSerological identification species groupspecies groupss
Virulence Virulence identificationidentification
detect endotoxin or exotoxindetect endotoxin or exotoxin
animal testanimal test rabbit guinea pirabbit guinea pigg
Susceptibility Susceptibility testtest
Slip methodSlip method determine the naturedetermine the nature
Test tube methodTest tube method quantitationquantitation(( detect MBCdetect MBC MICMIC ))
33 detect Agdetect Ag
mdashmdash mdashmdashserological identificationserological identification
highly specificityhighly specificity
sensitivity sensitivity
44 other detectionother detection
Detect metabolic productDetect metabolic product
bull bull PCR detect DNAPCR detect DNA
caution avoid contaminationcaution avoid contamination (faulse positive)(faulse positive)
bull bull DNA chips techniqueDNA chips technique
22 serological diagnosisserological diagnosis
-----detect Ab with known Ag-----detect Ab with known Ag
------specimens------specimens serumserum
------paired serum (acute------paired serum (acute convalescent serum)convalescent serum)
Compared acute serum and convalescenCompared acute serum and convalescent serum quantitativelyt serum quantitatively
A 4-fold or greater increase in Ab titerA 4-fold or greater increase in Ab titer su supports a diagnosis of recent infectionpports a diagnosis of recent infection
Serum containing a high titer of Abs of thSerum containing a high titer of Abs of thee IgM IgM subclass would suggest a subclass would suggest a current icurrent infectionnfection
commonly used methodscommonly used methods
Agglutination testAgglutination test Widal test---diagnose typhus and Widal test---diagnose typhus and
paratyphoidparatyphoid
Well-Felix---diagnose rickettsiosisWell-Felix---diagnose rickettsiosis
10486801048680
Neutralization testNeutralization test antildquoOrdquo test---diagnose rheumatic antildquoOrdquo test---diagnose rheumatic
diseasedisease
Section 2 virology diagnosisSection 2 virology diagnosis
ViralViral infeinfectionction
Electron microscopeElectron microscope
Light microscope inLight microscope inclusion bodyclusion body
isolationisolation
VirusVirus particleparticle
V AgV Ag
V nucleic acidV nucleic acid
Viral enzymeViral enzyme retroviridaseretroviridase
DetectDetect virusvirus
DetectDetect AbAb
Choose the appropriate specimenChoose the appropriate specimen
Aseptic operationAseptic operation add antibiotics to speciadd antibiotics to speci
menmen
Take it in acute phase Take it in acute phase
Low temperature conservationLow temperature conservation transport it transport it
promptlypromptly preserve in 50preserve in 50 glycerineglycerine
Serologial diagnose paired serumSerologial diagnose paired serum
Specimen Selection Collection and ProcessingSpecimen Selection Collection and Processing
二二 viral isolation and identifyingviral isolation and identifying
specimenspecimenAnimal inoculaAnimal inoculationtion
Embryonated eEmbryonated egggg
Cell cultureCell culture
positivepositive
negativenegative
identifyingidentifying
BlindBlind passagepassage22 - 3generatiogenerationn
negativenegative
Virus negatiVirus negativeve
asepsisasepsis
(一)(一) viral isolationviral isolation
11 animal inoculationanimal inoculation
chimpanzeemonkeyrabbitratchimpanzeemonkeyrabbitrat
22 embryonated egg inoculationembryonated egg inoculation
3 Cell culture3 Cell culture (( commonly usedcommonly used ))Primary and secondary cell culturePrimary and secondary cell culture
general proceduregeneral procedure
tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell
cell monolayercell monolayer
brokenbroken proteaseprotease
(primary cell)(primary cell)Serial subcultivationSerial subcultivation
Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)
features of virus cell culturefeatures of virus cell culture
A sensitive to many kinds of virusesA sensitive to many kinds of viruses
B cost highB cost high
C carry latent virusC carry latent virus
(2)Diploid(2)Diploid cellcell cultureculture
usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine
(3) secondary cell culture(3) secondary cell culture
sensitive to sensitive to multiplemultiple viruses viruses
high reproductive capacity long genera high reproductive capacity long generation timetion time
have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine
(( 二二 )) indexindex for reproduction of virusfor reproduction of virus
(1)Virus1)Virus infected cellinfected cell
(( cell roundinggathercell roundinggather
necrosisnecrosis ))
1 Cytopathic effect CPE)1 Cytopathic effect CPE)
CPECPE
(( 22 )) cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation
(( 33 )) viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell
CPECPE
22 hemadsorption hemadsorption HAd HAd
hemagglutininhemagglutinin (( HAHA )) on membrane of vion membrane of virus infected cell rus infected cell
Can Can adsorbadsorb vertebrate vertebrate RBCRBC
3 Interference3 Interference
4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange
( 三 ) viral quantitative assayHemagglutination test total amount
Plaque bacteriophage determination
ID50 or TCID50
Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le
sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex
cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a
nd hyphand hypha
22 cultureculture
specimen sabouraud mediumspecimen sabouraud medium
macroculture microculturemacroculture microculture
colonycolony
biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique
sporespore
3animal test3animal test
4skin test hypersensitivy4skin test hypersensitivy
(三)(三) mycology rapid diagnosismycology rapid diagnosis
serological test detect Ag or Abserological test detect Ag or Ab
detect nucleic acid of fungusdetect nucleic acid of fungus
detect mycotoxindetect mycotoxin
questionsquestions
11 definitiondefinition
serological identificationserological identification
and diagnosis)and diagnosis)
Cytopathic effect CPECytopathic effect CPE
hemadsorption HAdhemadsorption HAd
interferenceinterference
22 principle of Widalrsquos testprinciple of Widalrsquos test
33 principle of Gramrsquos stainingprinciple of Gramrsquos staining
Section 4Section 4
artificial active artificial active immunityimmunity
Artificial active immunityArtificial active immunity
stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid
vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA
A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow
th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni
citycity are repeatedly selected are repeatedly selected BCGBCG
The mutation is expressed only The mutation is expressed only
under certain conditionsunder certain conditions
may be may be useful in vaccinesuseful in vaccines
Conditional-lethal mutationsConditional-lethal mutations
temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation
the organisms can the organisms can replicate replicate at a at a
relativelyrelatively low low permissive temperature permissive temperature
but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive
temperaturetemperature
Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize
d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost
errdquo injectionerrdquo injection
A Live (attenuated) A Live (attenuated) vaccinesvaccines
B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate
responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe
toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves
artificial passive artificial passive immunization immunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
section 1 bacterial diagnosissection 1 bacterial diagnosis
bacteriological diagnosisbacteriological diagnosischeck pathogen and their componentscheck pathogen and their components
(( Ag products nucleic acidAg products nucleic acid )) detect Ag with known Abmdash detect Ag with known Abmdash serological identificationserological identificationdetect Ab in serum detect Ab in serum detect unknown Ab with known Agmdashdetect unknown Ab with known Agmdash serodiagnosisserodiagnosis
Steps precede the laboratory Steps precede the laboratory workwork
11 Choose the appropriate specimenChoose the appropriate specimen
22 Obtain the specimen avoid contaminatedObtain the specimen avoid contaminated
33 Transport the specimen promptly or Transport the specimen promptly or store it correctlystore it correctly
44 Provide essential informationProvide essential information
(二)(二) detection of pathogenic bacteria detection of pathogenic bacteria
11 morphology and structuremorphology and structure
22 isolating cultureisolating culture
33 detection of Agdetection of Ag
44 other methodsother methods
11 morphosis detectionmorphosis detection
procedureprocedure smear fix stain observe under smear fix stain observe under
microscopemicroscope
(( shapesizearrangement staining shapesizearrangement staining special structurespecial structure ) ) identify bacteriaidentify bacteria
major technique for detection of pathogmajor technique for detection of pathogenic bacteriaenic bacteria
(( 11 )) microscopemicroscope
Alight microscopeAlight microscope
resolutionresolution 025μm025μm
magnificationmagnification times1000times1000
B electronmicroscopeB electronmicroscope
resolutionresolution 1nm
magnification times100000
--cannot observe viable organism--cannot observe viable organism
C Other microscopeC Other microscope
bullbulldarkfield microscopedarkfield microscope
bull bullcontrast phase microscopecontrast phase microscope
bull bullfluorescence microscopefluorescence microscope
bull bullconfocal microscopeconfocal microscope
(( 22 )) staining techniquestaining technique
simple stainingsimple staining only use one dye only use one dye observe morphology arrangemen observe morphology arrangementt
compound stainingcompound staining use 2 kinds dyes or use 2 kinds dyes or more to identify bacteria more to identify bacteria
Gram stainingGram staining positivepositive (( violetviolet )) negativenegative (( redred ))Acid-fast stainingAcid-fast staining positivepositive (( redred )) negative negative (( blueblue ))Special stainingSpecial staining for bacterial capsule for bacterial capsule
spore flagella and so onspore flagella and so on
22 isolating culture and identifyisolating culture and identifyGeneral procedureGeneral procedure
specimen
Plain agar
Blood agar
Selective medium
cultural feature
nutrition
growing condition
Colony feature
hemolysis
pure
inoculate
stain initial identify (morphology)
Biochemical reaction
Serological identificationSerological identification species groupspecies groupss
Virulence Virulence identificationidentification
detect endotoxin or exotoxindetect endotoxin or exotoxin
animal testanimal test rabbit guinea pirabbit guinea pigg
Susceptibility Susceptibility testtest
Slip methodSlip method determine the naturedetermine the nature
Test tube methodTest tube method quantitationquantitation(( detect MBCdetect MBC MICMIC ))
33 detect Agdetect Ag
mdashmdash mdashmdashserological identificationserological identification
highly specificityhighly specificity
sensitivity sensitivity
44 other detectionother detection
Detect metabolic productDetect metabolic product
bull bull PCR detect DNAPCR detect DNA
caution avoid contaminationcaution avoid contamination (faulse positive)(faulse positive)
bull bull DNA chips techniqueDNA chips technique
22 serological diagnosisserological diagnosis
-----detect Ab with known Ag-----detect Ab with known Ag
------specimens------specimens serumserum
------paired serum (acute------paired serum (acute convalescent serum)convalescent serum)
Compared acute serum and convalescenCompared acute serum and convalescent serum quantitativelyt serum quantitatively
A 4-fold or greater increase in Ab titerA 4-fold or greater increase in Ab titer su supports a diagnosis of recent infectionpports a diagnosis of recent infection
Serum containing a high titer of Abs of thSerum containing a high titer of Abs of thee IgM IgM subclass would suggest a subclass would suggest a current icurrent infectionnfection
commonly used methodscommonly used methods
Agglutination testAgglutination test Widal test---diagnose typhus and Widal test---diagnose typhus and
paratyphoidparatyphoid
Well-Felix---diagnose rickettsiosisWell-Felix---diagnose rickettsiosis
10486801048680
Neutralization testNeutralization test antildquoOrdquo test---diagnose rheumatic antildquoOrdquo test---diagnose rheumatic
diseasedisease
Section 2 virology diagnosisSection 2 virology diagnosis
ViralViral infeinfectionction
Electron microscopeElectron microscope
Light microscope inLight microscope inclusion bodyclusion body
isolationisolation
VirusVirus particleparticle
V AgV Ag
V nucleic acidV nucleic acid
Viral enzymeViral enzyme retroviridaseretroviridase
DetectDetect virusvirus
DetectDetect AbAb
Choose the appropriate specimenChoose the appropriate specimen
Aseptic operationAseptic operation add antibiotics to speciadd antibiotics to speci
menmen
Take it in acute phase Take it in acute phase
Low temperature conservationLow temperature conservation transport it transport it
promptlypromptly preserve in 50preserve in 50 glycerineglycerine
Serologial diagnose paired serumSerologial diagnose paired serum
Specimen Selection Collection and ProcessingSpecimen Selection Collection and Processing
二二 viral isolation and identifyingviral isolation and identifying
specimenspecimenAnimal inoculaAnimal inoculationtion
Embryonated eEmbryonated egggg
Cell cultureCell culture
positivepositive
negativenegative
identifyingidentifying
BlindBlind passagepassage22 - 3generatiogenerationn
negativenegative
Virus negatiVirus negativeve
asepsisasepsis
(一)(一) viral isolationviral isolation
11 animal inoculationanimal inoculation
chimpanzeemonkeyrabbitratchimpanzeemonkeyrabbitrat
22 embryonated egg inoculationembryonated egg inoculation
3 Cell culture3 Cell culture (( commonly usedcommonly used ))Primary and secondary cell culturePrimary and secondary cell culture
general proceduregeneral procedure
tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell
cell monolayercell monolayer
brokenbroken proteaseprotease
(primary cell)(primary cell)Serial subcultivationSerial subcultivation
Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)
features of virus cell culturefeatures of virus cell culture
A sensitive to many kinds of virusesA sensitive to many kinds of viruses
B cost highB cost high
C carry latent virusC carry latent virus
(2)Diploid(2)Diploid cellcell cultureculture
usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine
(3) secondary cell culture(3) secondary cell culture
sensitive to sensitive to multiplemultiple viruses viruses
high reproductive capacity long genera high reproductive capacity long generation timetion time
have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine
(( 二二 )) indexindex for reproduction of virusfor reproduction of virus
(1)Virus1)Virus infected cellinfected cell
(( cell roundinggathercell roundinggather
necrosisnecrosis ))
1 Cytopathic effect CPE)1 Cytopathic effect CPE)
CPECPE
(( 22 )) cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation
(( 33 )) viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell
CPECPE
22 hemadsorption hemadsorption HAd HAd
hemagglutininhemagglutinin (( HAHA )) on membrane of vion membrane of virus infected cell rus infected cell
Can Can adsorbadsorb vertebrate vertebrate RBCRBC
3 Interference3 Interference
4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange
( 三 ) viral quantitative assayHemagglutination test total amount
Plaque bacteriophage determination
ID50 or TCID50
Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le
sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex
cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a
nd hyphand hypha
22 cultureculture
specimen sabouraud mediumspecimen sabouraud medium
macroculture microculturemacroculture microculture
colonycolony
biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique
sporespore
3animal test3animal test
4skin test hypersensitivy4skin test hypersensitivy
(三)(三) mycology rapid diagnosismycology rapid diagnosis
serological test detect Ag or Abserological test detect Ag or Ab
detect nucleic acid of fungusdetect nucleic acid of fungus
detect mycotoxindetect mycotoxin
questionsquestions
11 definitiondefinition
serological identificationserological identification
and diagnosis)and diagnosis)
Cytopathic effect CPECytopathic effect CPE
hemadsorption HAdhemadsorption HAd
interferenceinterference
22 principle of Widalrsquos testprinciple of Widalrsquos test
33 principle of Gramrsquos stainingprinciple of Gramrsquos staining
Section 4Section 4
artificial active artificial active immunityimmunity
Artificial active immunityArtificial active immunity
stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid
vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA
A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow
th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni
citycity are repeatedly selected are repeatedly selected BCGBCG
The mutation is expressed only The mutation is expressed only
under certain conditionsunder certain conditions
may be may be useful in vaccinesuseful in vaccines
Conditional-lethal mutationsConditional-lethal mutations
temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation
the organisms can the organisms can replicate replicate at a at a
relativelyrelatively low low permissive temperature permissive temperature
but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive
temperaturetemperature
Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize
d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost
errdquo injectionerrdquo injection
A Live (attenuated) A Live (attenuated) vaccinesvaccines
B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate
responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe
toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves
artificial passive artificial passive immunization immunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
Steps precede the laboratory Steps precede the laboratory workwork
11 Choose the appropriate specimenChoose the appropriate specimen
22 Obtain the specimen avoid contaminatedObtain the specimen avoid contaminated
33 Transport the specimen promptly or Transport the specimen promptly or store it correctlystore it correctly
44 Provide essential informationProvide essential information
(二)(二) detection of pathogenic bacteria detection of pathogenic bacteria
11 morphology and structuremorphology and structure
22 isolating cultureisolating culture
33 detection of Agdetection of Ag
44 other methodsother methods
11 morphosis detectionmorphosis detection
procedureprocedure smear fix stain observe under smear fix stain observe under
microscopemicroscope
(( shapesizearrangement staining shapesizearrangement staining special structurespecial structure ) ) identify bacteriaidentify bacteria
major technique for detection of pathogmajor technique for detection of pathogenic bacteriaenic bacteria
(( 11 )) microscopemicroscope
Alight microscopeAlight microscope
resolutionresolution 025μm025μm
magnificationmagnification times1000times1000
B electronmicroscopeB electronmicroscope
resolutionresolution 1nm
magnification times100000
--cannot observe viable organism--cannot observe viable organism
C Other microscopeC Other microscope
bullbulldarkfield microscopedarkfield microscope
bull bullcontrast phase microscopecontrast phase microscope
bull bullfluorescence microscopefluorescence microscope
bull bullconfocal microscopeconfocal microscope
(( 22 )) staining techniquestaining technique
simple stainingsimple staining only use one dye only use one dye observe morphology arrangemen observe morphology arrangementt
compound stainingcompound staining use 2 kinds dyes or use 2 kinds dyes or more to identify bacteria more to identify bacteria
Gram stainingGram staining positivepositive (( violetviolet )) negativenegative (( redred ))Acid-fast stainingAcid-fast staining positivepositive (( redred )) negative negative (( blueblue ))Special stainingSpecial staining for bacterial capsule for bacterial capsule
spore flagella and so onspore flagella and so on
22 isolating culture and identifyisolating culture and identifyGeneral procedureGeneral procedure
specimen
Plain agar
Blood agar
Selective medium
cultural feature
nutrition
growing condition
Colony feature
hemolysis
pure
inoculate
stain initial identify (morphology)
Biochemical reaction
Serological identificationSerological identification species groupspecies groupss
Virulence Virulence identificationidentification
detect endotoxin or exotoxindetect endotoxin or exotoxin
animal testanimal test rabbit guinea pirabbit guinea pigg
Susceptibility Susceptibility testtest
Slip methodSlip method determine the naturedetermine the nature
Test tube methodTest tube method quantitationquantitation(( detect MBCdetect MBC MICMIC ))
33 detect Agdetect Ag
mdashmdash mdashmdashserological identificationserological identification
highly specificityhighly specificity
sensitivity sensitivity
44 other detectionother detection
Detect metabolic productDetect metabolic product
bull bull PCR detect DNAPCR detect DNA
caution avoid contaminationcaution avoid contamination (faulse positive)(faulse positive)
bull bull DNA chips techniqueDNA chips technique
22 serological diagnosisserological diagnosis
-----detect Ab with known Ag-----detect Ab with known Ag
------specimens------specimens serumserum
------paired serum (acute------paired serum (acute convalescent serum)convalescent serum)
Compared acute serum and convalescenCompared acute serum and convalescent serum quantitativelyt serum quantitatively
A 4-fold or greater increase in Ab titerA 4-fold or greater increase in Ab titer su supports a diagnosis of recent infectionpports a diagnosis of recent infection
Serum containing a high titer of Abs of thSerum containing a high titer of Abs of thee IgM IgM subclass would suggest a subclass would suggest a current icurrent infectionnfection
commonly used methodscommonly used methods
Agglutination testAgglutination test Widal test---diagnose typhus and Widal test---diagnose typhus and
paratyphoidparatyphoid
Well-Felix---diagnose rickettsiosisWell-Felix---diagnose rickettsiosis
10486801048680
Neutralization testNeutralization test antildquoOrdquo test---diagnose rheumatic antildquoOrdquo test---diagnose rheumatic
diseasedisease
Section 2 virology diagnosisSection 2 virology diagnosis
ViralViral infeinfectionction
Electron microscopeElectron microscope
Light microscope inLight microscope inclusion bodyclusion body
isolationisolation
VirusVirus particleparticle
V AgV Ag
V nucleic acidV nucleic acid
Viral enzymeViral enzyme retroviridaseretroviridase
DetectDetect virusvirus
DetectDetect AbAb
Choose the appropriate specimenChoose the appropriate specimen
Aseptic operationAseptic operation add antibiotics to speciadd antibiotics to speci
menmen
Take it in acute phase Take it in acute phase
Low temperature conservationLow temperature conservation transport it transport it
promptlypromptly preserve in 50preserve in 50 glycerineglycerine
Serologial diagnose paired serumSerologial diagnose paired serum
Specimen Selection Collection and ProcessingSpecimen Selection Collection and Processing
二二 viral isolation and identifyingviral isolation and identifying
specimenspecimenAnimal inoculaAnimal inoculationtion
Embryonated eEmbryonated egggg
Cell cultureCell culture
positivepositive
negativenegative
identifyingidentifying
BlindBlind passagepassage22 - 3generatiogenerationn
negativenegative
Virus negatiVirus negativeve
asepsisasepsis
(一)(一) viral isolationviral isolation
11 animal inoculationanimal inoculation
chimpanzeemonkeyrabbitratchimpanzeemonkeyrabbitrat
22 embryonated egg inoculationembryonated egg inoculation
3 Cell culture3 Cell culture (( commonly usedcommonly used ))Primary and secondary cell culturePrimary and secondary cell culture
general proceduregeneral procedure
tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell
cell monolayercell monolayer
brokenbroken proteaseprotease
(primary cell)(primary cell)Serial subcultivationSerial subcultivation
Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)
features of virus cell culturefeatures of virus cell culture
A sensitive to many kinds of virusesA sensitive to many kinds of viruses
B cost highB cost high
C carry latent virusC carry latent virus
(2)Diploid(2)Diploid cellcell cultureculture
usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine
(3) secondary cell culture(3) secondary cell culture
sensitive to sensitive to multiplemultiple viruses viruses
high reproductive capacity long genera high reproductive capacity long generation timetion time
have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine
(( 二二 )) indexindex for reproduction of virusfor reproduction of virus
(1)Virus1)Virus infected cellinfected cell
(( cell roundinggathercell roundinggather
necrosisnecrosis ))
1 Cytopathic effect CPE)1 Cytopathic effect CPE)
CPECPE
(( 22 )) cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation
(( 33 )) viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell
CPECPE
22 hemadsorption hemadsorption HAd HAd
hemagglutininhemagglutinin (( HAHA )) on membrane of vion membrane of virus infected cell rus infected cell
Can Can adsorbadsorb vertebrate vertebrate RBCRBC
3 Interference3 Interference
4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange
( 三 ) viral quantitative assayHemagglutination test total amount
Plaque bacteriophage determination
ID50 or TCID50
Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le
sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex
cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a
nd hyphand hypha
22 cultureculture
specimen sabouraud mediumspecimen sabouraud medium
macroculture microculturemacroculture microculture
colonycolony
biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique
sporespore
3animal test3animal test
4skin test hypersensitivy4skin test hypersensitivy
(三)(三) mycology rapid diagnosismycology rapid diagnosis
serological test detect Ag or Abserological test detect Ag or Ab
detect nucleic acid of fungusdetect nucleic acid of fungus
detect mycotoxindetect mycotoxin
questionsquestions
11 definitiondefinition
serological identificationserological identification
and diagnosis)and diagnosis)
Cytopathic effect CPECytopathic effect CPE
hemadsorption HAdhemadsorption HAd
interferenceinterference
22 principle of Widalrsquos testprinciple of Widalrsquos test
33 principle of Gramrsquos stainingprinciple of Gramrsquos staining
Section 4Section 4
artificial active artificial active immunityimmunity
Artificial active immunityArtificial active immunity
stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid
vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA
A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow
th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni
citycity are repeatedly selected are repeatedly selected BCGBCG
The mutation is expressed only The mutation is expressed only
under certain conditionsunder certain conditions
may be may be useful in vaccinesuseful in vaccines
Conditional-lethal mutationsConditional-lethal mutations
temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation
the organisms can the organisms can replicate replicate at a at a
relativelyrelatively low low permissive temperature permissive temperature
but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive
temperaturetemperature
Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize
d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost
errdquo injectionerrdquo injection
A Live (attenuated) A Live (attenuated) vaccinesvaccines
B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate
responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe
toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves
artificial passive artificial passive immunization immunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
(二)(二) detection of pathogenic bacteria detection of pathogenic bacteria
11 morphology and structuremorphology and structure
22 isolating cultureisolating culture
33 detection of Agdetection of Ag
44 other methodsother methods
11 morphosis detectionmorphosis detection
procedureprocedure smear fix stain observe under smear fix stain observe under
microscopemicroscope
(( shapesizearrangement staining shapesizearrangement staining special structurespecial structure ) ) identify bacteriaidentify bacteria
major technique for detection of pathogmajor technique for detection of pathogenic bacteriaenic bacteria
(( 11 )) microscopemicroscope
Alight microscopeAlight microscope
resolutionresolution 025μm025μm
magnificationmagnification times1000times1000
B electronmicroscopeB electronmicroscope
resolutionresolution 1nm
magnification times100000
--cannot observe viable organism--cannot observe viable organism
C Other microscopeC Other microscope
bullbulldarkfield microscopedarkfield microscope
bull bullcontrast phase microscopecontrast phase microscope
bull bullfluorescence microscopefluorescence microscope
bull bullconfocal microscopeconfocal microscope
(( 22 )) staining techniquestaining technique
simple stainingsimple staining only use one dye only use one dye observe morphology arrangemen observe morphology arrangementt
compound stainingcompound staining use 2 kinds dyes or use 2 kinds dyes or more to identify bacteria more to identify bacteria
Gram stainingGram staining positivepositive (( violetviolet )) negativenegative (( redred ))Acid-fast stainingAcid-fast staining positivepositive (( redred )) negative negative (( blueblue ))Special stainingSpecial staining for bacterial capsule for bacterial capsule
spore flagella and so onspore flagella and so on
22 isolating culture and identifyisolating culture and identifyGeneral procedureGeneral procedure
specimen
Plain agar
Blood agar
Selective medium
cultural feature
nutrition
growing condition
Colony feature
hemolysis
pure
inoculate
stain initial identify (morphology)
Biochemical reaction
Serological identificationSerological identification species groupspecies groupss
Virulence Virulence identificationidentification
detect endotoxin or exotoxindetect endotoxin or exotoxin
animal testanimal test rabbit guinea pirabbit guinea pigg
Susceptibility Susceptibility testtest
Slip methodSlip method determine the naturedetermine the nature
Test tube methodTest tube method quantitationquantitation(( detect MBCdetect MBC MICMIC ))
33 detect Agdetect Ag
mdashmdash mdashmdashserological identificationserological identification
highly specificityhighly specificity
sensitivity sensitivity
44 other detectionother detection
Detect metabolic productDetect metabolic product
bull bull PCR detect DNAPCR detect DNA
caution avoid contaminationcaution avoid contamination (faulse positive)(faulse positive)
bull bull DNA chips techniqueDNA chips technique
22 serological diagnosisserological diagnosis
-----detect Ab with known Ag-----detect Ab with known Ag
------specimens------specimens serumserum
------paired serum (acute------paired serum (acute convalescent serum)convalescent serum)
Compared acute serum and convalescenCompared acute serum and convalescent serum quantitativelyt serum quantitatively
A 4-fold or greater increase in Ab titerA 4-fold or greater increase in Ab titer su supports a diagnosis of recent infectionpports a diagnosis of recent infection
Serum containing a high titer of Abs of thSerum containing a high titer of Abs of thee IgM IgM subclass would suggest a subclass would suggest a current icurrent infectionnfection
commonly used methodscommonly used methods
Agglutination testAgglutination test Widal test---diagnose typhus and Widal test---diagnose typhus and
paratyphoidparatyphoid
Well-Felix---diagnose rickettsiosisWell-Felix---diagnose rickettsiosis
10486801048680
Neutralization testNeutralization test antildquoOrdquo test---diagnose rheumatic antildquoOrdquo test---diagnose rheumatic
diseasedisease
Section 2 virology diagnosisSection 2 virology diagnosis
ViralViral infeinfectionction
Electron microscopeElectron microscope
Light microscope inLight microscope inclusion bodyclusion body
isolationisolation
VirusVirus particleparticle
V AgV Ag
V nucleic acidV nucleic acid
Viral enzymeViral enzyme retroviridaseretroviridase
DetectDetect virusvirus
DetectDetect AbAb
Choose the appropriate specimenChoose the appropriate specimen
Aseptic operationAseptic operation add antibiotics to speciadd antibiotics to speci
menmen
Take it in acute phase Take it in acute phase
Low temperature conservationLow temperature conservation transport it transport it
promptlypromptly preserve in 50preserve in 50 glycerineglycerine
Serologial diagnose paired serumSerologial diagnose paired serum
Specimen Selection Collection and ProcessingSpecimen Selection Collection and Processing
二二 viral isolation and identifyingviral isolation and identifying
specimenspecimenAnimal inoculaAnimal inoculationtion
Embryonated eEmbryonated egggg
Cell cultureCell culture
positivepositive
negativenegative
identifyingidentifying
BlindBlind passagepassage22 - 3generatiogenerationn
negativenegative
Virus negatiVirus negativeve
asepsisasepsis
(一)(一) viral isolationviral isolation
11 animal inoculationanimal inoculation
chimpanzeemonkeyrabbitratchimpanzeemonkeyrabbitrat
22 embryonated egg inoculationembryonated egg inoculation
3 Cell culture3 Cell culture (( commonly usedcommonly used ))Primary and secondary cell culturePrimary and secondary cell culture
general proceduregeneral procedure
tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell
cell monolayercell monolayer
brokenbroken proteaseprotease
(primary cell)(primary cell)Serial subcultivationSerial subcultivation
Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)
features of virus cell culturefeatures of virus cell culture
A sensitive to many kinds of virusesA sensitive to many kinds of viruses
B cost highB cost high
C carry latent virusC carry latent virus
(2)Diploid(2)Diploid cellcell cultureculture
usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine
(3) secondary cell culture(3) secondary cell culture
sensitive to sensitive to multiplemultiple viruses viruses
high reproductive capacity long genera high reproductive capacity long generation timetion time
have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine
(( 二二 )) indexindex for reproduction of virusfor reproduction of virus
(1)Virus1)Virus infected cellinfected cell
(( cell roundinggathercell roundinggather
necrosisnecrosis ))
1 Cytopathic effect CPE)1 Cytopathic effect CPE)
CPECPE
(( 22 )) cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation
(( 33 )) viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell
CPECPE
22 hemadsorption hemadsorption HAd HAd
hemagglutininhemagglutinin (( HAHA )) on membrane of vion membrane of virus infected cell rus infected cell
Can Can adsorbadsorb vertebrate vertebrate RBCRBC
3 Interference3 Interference
4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange
( 三 ) viral quantitative assayHemagglutination test total amount
Plaque bacteriophage determination
ID50 or TCID50
Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le
sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex
cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a
nd hyphand hypha
22 cultureculture
specimen sabouraud mediumspecimen sabouraud medium
macroculture microculturemacroculture microculture
colonycolony
biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique
sporespore
3animal test3animal test
4skin test hypersensitivy4skin test hypersensitivy
(三)(三) mycology rapid diagnosismycology rapid diagnosis
serological test detect Ag or Abserological test detect Ag or Ab
detect nucleic acid of fungusdetect nucleic acid of fungus
detect mycotoxindetect mycotoxin
questionsquestions
11 definitiondefinition
serological identificationserological identification
and diagnosis)and diagnosis)
Cytopathic effect CPECytopathic effect CPE
hemadsorption HAdhemadsorption HAd
interferenceinterference
22 principle of Widalrsquos testprinciple of Widalrsquos test
33 principle of Gramrsquos stainingprinciple of Gramrsquos staining
Section 4Section 4
artificial active artificial active immunityimmunity
Artificial active immunityArtificial active immunity
stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid
vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA
A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow
th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni
citycity are repeatedly selected are repeatedly selected BCGBCG
The mutation is expressed only The mutation is expressed only
under certain conditionsunder certain conditions
may be may be useful in vaccinesuseful in vaccines
Conditional-lethal mutationsConditional-lethal mutations
temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation
the organisms can the organisms can replicate replicate at a at a
relativelyrelatively low low permissive temperature permissive temperature
but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive
temperaturetemperature
Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize
d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost
errdquo injectionerrdquo injection
A Live (attenuated) A Live (attenuated) vaccinesvaccines
B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate
responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe
toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves
artificial passive artificial passive immunization immunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
11 morphosis detectionmorphosis detection
procedureprocedure smear fix stain observe under smear fix stain observe under
microscopemicroscope
(( shapesizearrangement staining shapesizearrangement staining special structurespecial structure ) ) identify bacteriaidentify bacteria
major technique for detection of pathogmajor technique for detection of pathogenic bacteriaenic bacteria
(( 11 )) microscopemicroscope
Alight microscopeAlight microscope
resolutionresolution 025μm025μm
magnificationmagnification times1000times1000
B electronmicroscopeB electronmicroscope
resolutionresolution 1nm
magnification times100000
--cannot observe viable organism--cannot observe viable organism
C Other microscopeC Other microscope
bullbulldarkfield microscopedarkfield microscope
bull bullcontrast phase microscopecontrast phase microscope
bull bullfluorescence microscopefluorescence microscope
bull bullconfocal microscopeconfocal microscope
(( 22 )) staining techniquestaining technique
simple stainingsimple staining only use one dye only use one dye observe morphology arrangemen observe morphology arrangementt
compound stainingcompound staining use 2 kinds dyes or use 2 kinds dyes or more to identify bacteria more to identify bacteria
Gram stainingGram staining positivepositive (( violetviolet )) negativenegative (( redred ))Acid-fast stainingAcid-fast staining positivepositive (( redred )) negative negative (( blueblue ))Special stainingSpecial staining for bacterial capsule for bacterial capsule
spore flagella and so onspore flagella and so on
22 isolating culture and identifyisolating culture and identifyGeneral procedureGeneral procedure
specimen
Plain agar
Blood agar
Selective medium
cultural feature
nutrition
growing condition
Colony feature
hemolysis
pure
inoculate
stain initial identify (morphology)
Biochemical reaction
Serological identificationSerological identification species groupspecies groupss
Virulence Virulence identificationidentification
detect endotoxin or exotoxindetect endotoxin or exotoxin
animal testanimal test rabbit guinea pirabbit guinea pigg
Susceptibility Susceptibility testtest
Slip methodSlip method determine the naturedetermine the nature
Test tube methodTest tube method quantitationquantitation(( detect MBCdetect MBC MICMIC ))
33 detect Agdetect Ag
mdashmdash mdashmdashserological identificationserological identification
highly specificityhighly specificity
sensitivity sensitivity
44 other detectionother detection
Detect metabolic productDetect metabolic product
bull bull PCR detect DNAPCR detect DNA
caution avoid contaminationcaution avoid contamination (faulse positive)(faulse positive)
bull bull DNA chips techniqueDNA chips technique
22 serological diagnosisserological diagnosis
-----detect Ab with known Ag-----detect Ab with known Ag
------specimens------specimens serumserum
------paired serum (acute------paired serum (acute convalescent serum)convalescent serum)
Compared acute serum and convalescenCompared acute serum and convalescent serum quantitativelyt serum quantitatively
A 4-fold or greater increase in Ab titerA 4-fold or greater increase in Ab titer su supports a diagnosis of recent infectionpports a diagnosis of recent infection
Serum containing a high titer of Abs of thSerum containing a high titer of Abs of thee IgM IgM subclass would suggest a subclass would suggest a current icurrent infectionnfection
commonly used methodscommonly used methods
Agglutination testAgglutination test Widal test---diagnose typhus and Widal test---diagnose typhus and
paratyphoidparatyphoid
Well-Felix---diagnose rickettsiosisWell-Felix---diagnose rickettsiosis
10486801048680
Neutralization testNeutralization test antildquoOrdquo test---diagnose rheumatic antildquoOrdquo test---diagnose rheumatic
diseasedisease
Section 2 virology diagnosisSection 2 virology diagnosis
ViralViral infeinfectionction
Electron microscopeElectron microscope
Light microscope inLight microscope inclusion bodyclusion body
isolationisolation
VirusVirus particleparticle
V AgV Ag
V nucleic acidV nucleic acid
Viral enzymeViral enzyme retroviridaseretroviridase
DetectDetect virusvirus
DetectDetect AbAb
Choose the appropriate specimenChoose the appropriate specimen
Aseptic operationAseptic operation add antibiotics to speciadd antibiotics to speci
menmen
Take it in acute phase Take it in acute phase
Low temperature conservationLow temperature conservation transport it transport it
promptlypromptly preserve in 50preserve in 50 glycerineglycerine
Serologial diagnose paired serumSerologial diagnose paired serum
Specimen Selection Collection and ProcessingSpecimen Selection Collection and Processing
二二 viral isolation and identifyingviral isolation and identifying
specimenspecimenAnimal inoculaAnimal inoculationtion
Embryonated eEmbryonated egggg
Cell cultureCell culture
positivepositive
negativenegative
identifyingidentifying
BlindBlind passagepassage22 - 3generatiogenerationn
negativenegative
Virus negatiVirus negativeve
asepsisasepsis
(一)(一) viral isolationviral isolation
11 animal inoculationanimal inoculation
chimpanzeemonkeyrabbitratchimpanzeemonkeyrabbitrat
22 embryonated egg inoculationembryonated egg inoculation
3 Cell culture3 Cell culture (( commonly usedcommonly used ))Primary and secondary cell culturePrimary and secondary cell culture
general proceduregeneral procedure
tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell
cell monolayercell monolayer
brokenbroken proteaseprotease
(primary cell)(primary cell)Serial subcultivationSerial subcultivation
Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)
features of virus cell culturefeatures of virus cell culture
A sensitive to many kinds of virusesA sensitive to many kinds of viruses
B cost highB cost high
C carry latent virusC carry latent virus
(2)Diploid(2)Diploid cellcell cultureculture
usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine
(3) secondary cell culture(3) secondary cell culture
sensitive to sensitive to multiplemultiple viruses viruses
high reproductive capacity long genera high reproductive capacity long generation timetion time
have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine
(( 二二 )) indexindex for reproduction of virusfor reproduction of virus
(1)Virus1)Virus infected cellinfected cell
(( cell roundinggathercell roundinggather
necrosisnecrosis ))
1 Cytopathic effect CPE)1 Cytopathic effect CPE)
CPECPE
(( 22 )) cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation
(( 33 )) viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell
CPECPE
22 hemadsorption hemadsorption HAd HAd
hemagglutininhemagglutinin (( HAHA )) on membrane of vion membrane of virus infected cell rus infected cell
Can Can adsorbadsorb vertebrate vertebrate RBCRBC
3 Interference3 Interference
4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange
( 三 ) viral quantitative assayHemagglutination test total amount
Plaque bacteriophage determination
ID50 or TCID50
Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le
sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex
cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a
nd hyphand hypha
22 cultureculture
specimen sabouraud mediumspecimen sabouraud medium
macroculture microculturemacroculture microculture
colonycolony
biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique
sporespore
3animal test3animal test
4skin test hypersensitivy4skin test hypersensitivy
(三)(三) mycology rapid diagnosismycology rapid diagnosis
serological test detect Ag or Abserological test detect Ag or Ab
detect nucleic acid of fungusdetect nucleic acid of fungus
detect mycotoxindetect mycotoxin
questionsquestions
11 definitiondefinition
serological identificationserological identification
and diagnosis)and diagnosis)
Cytopathic effect CPECytopathic effect CPE
hemadsorption HAdhemadsorption HAd
interferenceinterference
22 principle of Widalrsquos testprinciple of Widalrsquos test
33 principle of Gramrsquos stainingprinciple of Gramrsquos staining
Section 4Section 4
artificial active artificial active immunityimmunity
Artificial active immunityArtificial active immunity
stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid
vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA
A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow
th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni
citycity are repeatedly selected are repeatedly selected BCGBCG
The mutation is expressed only The mutation is expressed only
under certain conditionsunder certain conditions
may be may be useful in vaccinesuseful in vaccines
Conditional-lethal mutationsConditional-lethal mutations
temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation
the organisms can the organisms can replicate replicate at a at a
relativelyrelatively low low permissive temperature permissive temperature
but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive
temperaturetemperature
Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize
d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost
errdquo injectionerrdquo injection
A Live (attenuated) A Live (attenuated) vaccinesvaccines
B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate
responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe
toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves
artificial passive artificial passive immunization immunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
major technique for detection of pathogmajor technique for detection of pathogenic bacteriaenic bacteria
(( 11 )) microscopemicroscope
Alight microscopeAlight microscope
resolutionresolution 025μm025μm
magnificationmagnification times1000times1000
B electronmicroscopeB electronmicroscope
resolutionresolution 1nm
magnification times100000
--cannot observe viable organism--cannot observe viable organism
C Other microscopeC Other microscope
bullbulldarkfield microscopedarkfield microscope
bull bullcontrast phase microscopecontrast phase microscope
bull bullfluorescence microscopefluorescence microscope
bull bullconfocal microscopeconfocal microscope
(( 22 )) staining techniquestaining technique
simple stainingsimple staining only use one dye only use one dye observe morphology arrangemen observe morphology arrangementt
compound stainingcompound staining use 2 kinds dyes or use 2 kinds dyes or more to identify bacteria more to identify bacteria
Gram stainingGram staining positivepositive (( violetviolet )) negativenegative (( redred ))Acid-fast stainingAcid-fast staining positivepositive (( redred )) negative negative (( blueblue ))Special stainingSpecial staining for bacterial capsule for bacterial capsule
spore flagella and so onspore flagella and so on
22 isolating culture and identifyisolating culture and identifyGeneral procedureGeneral procedure
specimen
Plain agar
Blood agar
Selective medium
cultural feature
nutrition
growing condition
Colony feature
hemolysis
pure
inoculate
stain initial identify (morphology)
Biochemical reaction
Serological identificationSerological identification species groupspecies groupss
Virulence Virulence identificationidentification
detect endotoxin or exotoxindetect endotoxin or exotoxin
animal testanimal test rabbit guinea pirabbit guinea pigg
Susceptibility Susceptibility testtest
Slip methodSlip method determine the naturedetermine the nature
Test tube methodTest tube method quantitationquantitation(( detect MBCdetect MBC MICMIC ))
33 detect Agdetect Ag
mdashmdash mdashmdashserological identificationserological identification
highly specificityhighly specificity
sensitivity sensitivity
44 other detectionother detection
Detect metabolic productDetect metabolic product
bull bull PCR detect DNAPCR detect DNA
caution avoid contaminationcaution avoid contamination (faulse positive)(faulse positive)
bull bull DNA chips techniqueDNA chips technique
22 serological diagnosisserological diagnosis
-----detect Ab with known Ag-----detect Ab with known Ag
------specimens------specimens serumserum
------paired serum (acute------paired serum (acute convalescent serum)convalescent serum)
Compared acute serum and convalescenCompared acute serum and convalescent serum quantitativelyt serum quantitatively
A 4-fold or greater increase in Ab titerA 4-fold or greater increase in Ab titer su supports a diagnosis of recent infectionpports a diagnosis of recent infection
Serum containing a high titer of Abs of thSerum containing a high titer of Abs of thee IgM IgM subclass would suggest a subclass would suggest a current icurrent infectionnfection
commonly used methodscommonly used methods
Agglutination testAgglutination test Widal test---diagnose typhus and Widal test---diagnose typhus and
paratyphoidparatyphoid
Well-Felix---diagnose rickettsiosisWell-Felix---diagnose rickettsiosis
10486801048680
Neutralization testNeutralization test antildquoOrdquo test---diagnose rheumatic antildquoOrdquo test---diagnose rheumatic
diseasedisease
Section 2 virology diagnosisSection 2 virology diagnosis
ViralViral infeinfectionction
Electron microscopeElectron microscope
Light microscope inLight microscope inclusion bodyclusion body
isolationisolation
VirusVirus particleparticle
V AgV Ag
V nucleic acidV nucleic acid
Viral enzymeViral enzyme retroviridaseretroviridase
DetectDetect virusvirus
DetectDetect AbAb
Choose the appropriate specimenChoose the appropriate specimen
Aseptic operationAseptic operation add antibiotics to speciadd antibiotics to speci
menmen
Take it in acute phase Take it in acute phase
Low temperature conservationLow temperature conservation transport it transport it
promptlypromptly preserve in 50preserve in 50 glycerineglycerine
Serologial diagnose paired serumSerologial diagnose paired serum
Specimen Selection Collection and ProcessingSpecimen Selection Collection and Processing
二二 viral isolation and identifyingviral isolation and identifying
specimenspecimenAnimal inoculaAnimal inoculationtion
Embryonated eEmbryonated egggg
Cell cultureCell culture
positivepositive
negativenegative
identifyingidentifying
BlindBlind passagepassage22 - 3generatiogenerationn
negativenegative
Virus negatiVirus negativeve
asepsisasepsis
(一)(一) viral isolationviral isolation
11 animal inoculationanimal inoculation
chimpanzeemonkeyrabbitratchimpanzeemonkeyrabbitrat
22 embryonated egg inoculationembryonated egg inoculation
3 Cell culture3 Cell culture (( commonly usedcommonly used ))Primary and secondary cell culturePrimary and secondary cell culture
general proceduregeneral procedure
tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell
cell monolayercell monolayer
brokenbroken proteaseprotease
(primary cell)(primary cell)Serial subcultivationSerial subcultivation
Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)
features of virus cell culturefeatures of virus cell culture
A sensitive to many kinds of virusesA sensitive to many kinds of viruses
B cost highB cost high
C carry latent virusC carry latent virus
(2)Diploid(2)Diploid cellcell cultureculture
usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine
(3) secondary cell culture(3) secondary cell culture
sensitive to sensitive to multiplemultiple viruses viruses
high reproductive capacity long genera high reproductive capacity long generation timetion time
have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine
(( 二二 )) indexindex for reproduction of virusfor reproduction of virus
(1)Virus1)Virus infected cellinfected cell
(( cell roundinggathercell roundinggather
necrosisnecrosis ))
1 Cytopathic effect CPE)1 Cytopathic effect CPE)
CPECPE
(( 22 )) cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation
(( 33 )) viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell
CPECPE
22 hemadsorption hemadsorption HAd HAd
hemagglutininhemagglutinin (( HAHA )) on membrane of vion membrane of virus infected cell rus infected cell
Can Can adsorbadsorb vertebrate vertebrate RBCRBC
3 Interference3 Interference
4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange
( 三 ) viral quantitative assayHemagglutination test total amount
Plaque bacteriophage determination
ID50 or TCID50
Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le
sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex
cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a
nd hyphand hypha
22 cultureculture
specimen sabouraud mediumspecimen sabouraud medium
macroculture microculturemacroculture microculture
colonycolony
biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique
sporespore
3animal test3animal test
4skin test hypersensitivy4skin test hypersensitivy
(三)(三) mycology rapid diagnosismycology rapid diagnosis
serological test detect Ag or Abserological test detect Ag or Ab
detect nucleic acid of fungusdetect nucleic acid of fungus
detect mycotoxindetect mycotoxin
questionsquestions
11 definitiondefinition
serological identificationserological identification
and diagnosis)and diagnosis)
Cytopathic effect CPECytopathic effect CPE
hemadsorption HAdhemadsorption HAd
interferenceinterference
22 principle of Widalrsquos testprinciple of Widalrsquos test
33 principle of Gramrsquos stainingprinciple of Gramrsquos staining
Section 4Section 4
artificial active artificial active immunityimmunity
Artificial active immunityArtificial active immunity
stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid
vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA
A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow
th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni
citycity are repeatedly selected are repeatedly selected BCGBCG
The mutation is expressed only The mutation is expressed only
under certain conditionsunder certain conditions
may be may be useful in vaccinesuseful in vaccines
Conditional-lethal mutationsConditional-lethal mutations
temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation
the organisms can the organisms can replicate replicate at a at a
relativelyrelatively low low permissive temperature permissive temperature
but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive
temperaturetemperature
Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize
d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost
errdquo injectionerrdquo injection
A Live (attenuated) A Live (attenuated) vaccinesvaccines
B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate
responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe
toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves
artificial passive artificial passive immunization immunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
B electronmicroscopeB electronmicroscope
resolutionresolution 1nm
magnification times100000
--cannot observe viable organism--cannot observe viable organism
C Other microscopeC Other microscope
bullbulldarkfield microscopedarkfield microscope
bull bullcontrast phase microscopecontrast phase microscope
bull bullfluorescence microscopefluorescence microscope
bull bullconfocal microscopeconfocal microscope
(( 22 )) staining techniquestaining technique
simple stainingsimple staining only use one dye only use one dye observe morphology arrangemen observe morphology arrangementt
compound stainingcompound staining use 2 kinds dyes or use 2 kinds dyes or more to identify bacteria more to identify bacteria
Gram stainingGram staining positivepositive (( violetviolet )) negativenegative (( redred ))Acid-fast stainingAcid-fast staining positivepositive (( redred )) negative negative (( blueblue ))Special stainingSpecial staining for bacterial capsule for bacterial capsule
spore flagella and so onspore flagella and so on
22 isolating culture and identifyisolating culture and identifyGeneral procedureGeneral procedure
specimen
Plain agar
Blood agar
Selective medium
cultural feature
nutrition
growing condition
Colony feature
hemolysis
pure
inoculate
stain initial identify (morphology)
Biochemical reaction
Serological identificationSerological identification species groupspecies groupss
Virulence Virulence identificationidentification
detect endotoxin or exotoxindetect endotoxin or exotoxin
animal testanimal test rabbit guinea pirabbit guinea pigg
Susceptibility Susceptibility testtest
Slip methodSlip method determine the naturedetermine the nature
Test tube methodTest tube method quantitationquantitation(( detect MBCdetect MBC MICMIC ))
33 detect Agdetect Ag
mdashmdash mdashmdashserological identificationserological identification
highly specificityhighly specificity
sensitivity sensitivity
44 other detectionother detection
Detect metabolic productDetect metabolic product
bull bull PCR detect DNAPCR detect DNA
caution avoid contaminationcaution avoid contamination (faulse positive)(faulse positive)
bull bull DNA chips techniqueDNA chips technique
22 serological diagnosisserological diagnosis
-----detect Ab with known Ag-----detect Ab with known Ag
------specimens------specimens serumserum
------paired serum (acute------paired serum (acute convalescent serum)convalescent serum)
Compared acute serum and convalescenCompared acute serum and convalescent serum quantitativelyt serum quantitatively
A 4-fold or greater increase in Ab titerA 4-fold or greater increase in Ab titer su supports a diagnosis of recent infectionpports a diagnosis of recent infection
Serum containing a high titer of Abs of thSerum containing a high titer of Abs of thee IgM IgM subclass would suggest a subclass would suggest a current icurrent infectionnfection
commonly used methodscommonly used methods
Agglutination testAgglutination test Widal test---diagnose typhus and Widal test---diagnose typhus and
paratyphoidparatyphoid
Well-Felix---diagnose rickettsiosisWell-Felix---diagnose rickettsiosis
10486801048680
Neutralization testNeutralization test antildquoOrdquo test---diagnose rheumatic antildquoOrdquo test---diagnose rheumatic
diseasedisease
Section 2 virology diagnosisSection 2 virology diagnosis
ViralViral infeinfectionction
Electron microscopeElectron microscope
Light microscope inLight microscope inclusion bodyclusion body
isolationisolation
VirusVirus particleparticle
V AgV Ag
V nucleic acidV nucleic acid
Viral enzymeViral enzyme retroviridaseretroviridase
DetectDetect virusvirus
DetectDetect AbAb
Choose the appropriate specimenChoose the appropriate specimen
Aseptic operationAseptic operation add antibiotics to speciadd antibiotics to speci
menmen
Take it in acute phase Take it in acute phase
Low temperature conservationLow temperature conservation transport it transport it
promptlypromptly preserve in 50preserve in 50 glycerineglycerine
Serologial diagnose paired serumSerologial diagnose paired serum
Specimen Selection Collection and ProcessingSpecimen Selection Collection and Processing
二二 viral isolation and identifyingviral isolation and identifying
specimenspecimenAnimal inoculaAnimal inoculationtion
Embryonated eEmbryonated egggg
Cell cultureCell culture
positivepositive
negativenegative
identifyingidentifying
BlindBlind passagepassage22 - 3generatiogenerationn
negativenegative
Virus negatiVirus negativeve
asepsisasepsis
(一)(一) viral isolationviral isolation
11 animal inoculationanimal inoculation
chimpanzeemonkeyrabbitratchimpanzeemonkeyrabbitrat
22 embryonated egg inoculationembryonated egg inoculation
3 Cell culture3 Cell culture (( commonly usedcommonly used ))Primary and secondary cell culturePrimary and secondary cell culture
general proceduregeneral procedure
tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell
cell monolayercell monolayer
brokenbroken proteaseprotease
(primary cell)(primary cell)Serial subcultivationSerial subcultivation
Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)
features of virus cell culturefeatures of virus cell culture
A sensitive to many kinds of virusesA sensitive to many kinds of viruses
B cost highB cost high
C carry latent virusC carry latent virus
(2)Diploid(2)Diploid cellcell cultureculture
usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine
(3) secondary cell culture(3) secondary cell culture
sensitive to sensitive to multiplemultiple viruses viruses
high reproductive capacity long genera high reproductive capacity long generation timetion time
have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine
(( 二二 )) indexindex for reproduction of virusfor reproduction of virus
(1)Virus1)Virus infected cellinfected cell
(( cell roundinggathercell roundinggather
necrosisnecrosis ))
1 Cytopathic effect CPE)1 Cytopathic effect CPE)
CPECPE
(( 22 )) cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation
(( 33 )) viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell
CPECPE
22 hemadsorption hemadsorption HAd HAd
hemagglutininhemagglutinin (( HAHA )) on membrane of vion membrane of virus infected cell rus infected cell
Can Can adsorbadsorb vertebrate vertebrate RBCRBC
3 Interference3 Interference
4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange
( 三 ) viral quantitative assayHemagglutination test total amount
Plaque bacteriophage determination
ID50 or TCID50
Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le
sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex
cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a
nd hyphand hypha
22 cultureculture
specimen sabouraud mediumspecimen sabouraud medium
macroculture microculturemacroculture microculture
colonycolony
biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique
sporespore
3animal test3animal test
4skin test hypersensitivy4skin test hypersensitivy
(三)(三) mycology rapid diagnosismycology rapid diagnosis
serological test detect Ag or Abserological test detect Ag or Ab
detect nucleic acid of fungusdetect nucleic acid of fungus
detect mycotoxindetect mycotoxin
questionsquestions
11 definitiondefinition
serological identificationserological identification
and diagnosis)and diagnosis)
Cytopathic effect CPECytopathic effect CPE
hemadsorption HAdhemadsorption HAd
interferenceinterference
22 principle of Widalrsquos testprinciple of Widalrsquos test
33 principle of Gramrsquos stainingprinciple of Gramrsquos staining
Section 4Section 4
artificial active artificial active immunityimmunity
Artificial active immunityArtificial active immunity
stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid
vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA
A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow
th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni
citycity are repeatedly selected are repeatedly selected BCGBCG
The mutation is expressed only The mutation is expressed only
under certain conditionsunder certain conditions
may be may be useful in vaccinesuseful in vaccines
Conditional-lethal mutationsConditional-lethal mutations
temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation
the organisms can the organisms can replicate replicate at a at a
relativelyrelatively low low permissive temperature permissive temperature
but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive
temperaturetemperature
Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize
d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost
errdquo injectionerrdquo injection
A Live (attenuated) A Live (attenuated) vaccinesvaccines
B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate
responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe
toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves
artificial passive artificial passive immunization immunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
C Other microscopeC Other microscope
bullbulldarkfield microscopedarkfield microscope
bull bullcontrast phase microscopecontrast phase microscope
bull bullfluorescence microscopefluorescence microscope
bull bullconfocal microscopeconfocal microscope
(( 22 )) staining techniquestaining technique
simple stainingsimple staining only use one dye only use one dye observe morphology arrangemen observe morphology arrangementt
compound stainingcompound staining use 2 kinds dyes or use 2 kinds dyes or more to identify bacteria more to identify bacteria
Gram stainingGram staining positivepositive (( violetviolet )) negativenegative (( redred ))Acid-fast stainingAcid-fast staining positivepositive (( redred )) negative negative (( blueblue ))Special stainingSpecial staining for bacterial capsule for bacterial capsule
spore flagella and so onspore flagella and so on
22 isolating culture and identifyisolating culture and identifyGeneral procedureGeneral procedure
specimen
Plain agar
Blood agar
Selective medium
cultural feature
nutrition
growing condition
Colony feature
hemolysis
pure
inoculate
stain initial identify (morphology)
Biochemical reaction
Serological identificationSerological identification species groupspecies groupss
Virulence Virulence identificationidentification
detect endotoxin or exotoxindetect endotoxin or exotoxin
animal testanimal test rabbit guinea pirabbit guinea pigg
Susceptibility Susceptibility testtest
Slip methodSlip method determine the naturedetermine the nature
Test tube methodTest tube method quantitationquantitation(( detect MBCdetect MBC MICMIC ))
33 detect Agdetect Ag
mdashmdash mdashmdashserological identificationserological identification
highly specificityhighly specificity
sensitivity sensitivity
44 other detectionother detection
Detect metabolic productDetect metabolic product
bull bull PCR detect DNAPCR detect DNA
caution avoid contaminationcaution avoid contamination (faulse positive)(faulse positive)
bull bull DNA chips techniqueDNA chips technique
22 serological diagnosisserological diagnosis
-----detect Ab with known Ag-----detect Ab with known Ag
------specimens------specimens serumserum
------paired serum (acute------paired serum (acute convalescent serum)convalescent serum)
Compared acute serum and convalescenCompared acute serum and convalescent serum quantitativelyt serum quantitatively
A 4-fold or greater increase in Ab titerA 4-fold or greater increase in Ab titer su supports a diagnosis of recent infectionpports a diagnosis of recent infection
Serum containing a high titer of Abs of thSerum containing a high titer of Abs of thee IgM IgM subclass would suggest a subclass would suggest a current icurrent infectionnfection
commonly used methodscommonly used methods
Agglutination testAgglutination test Widal test---diagnose typhus and Widal test---diagnose typhus and
paratyphoidparatyphoid
Well-Felix---diagnose rickettsiosisWell-Felix---diagnose rickettsiosis
10486801048680
Neutralization testNeutralization test antildquoOrdquo test---diagnose rheumatic antildquoOrdquo test---diagnose rheumatic
diseasedisease
Section 2 virology diagnosisSection 2 virology diagnosis
ViralViral infeinfectionction
Electron microscopeElectron microscope
Light microscope inLight microscope inclusion bodyclusion body
isolationisolation
VirusVirus particleparticle
V AgV Ag
V nucleic acidV nucleic acid
Viral enzymeViral enzyme retroviridaseretroviridase
DetectDetect virusvirus
DetectDetect AbAb
Choose the appropriate specimenChoose the appropriate specimen
Aseptic operationAseptic operation add antibiotics to speciadd antibiotics to speci
menmen
Take it in acute phase Take it in acute phase
Low temperature conservationLow temperature conservation transport it transport it
promptlypromptly preserve in 50preserve in 50 glycerineglycerine
Serologial diagnose paired serumSerologial diagnose paired serum
Specimen Selection Collection and ProcessingSpecimen Selection Collection and Processing
二二 viral isolation and identifyingviral isolation and identifying
specimenspecimenAnimal inoculaAnimal inoculationtion
Embryonated eEmbryonated egggg
Cell cultureCell culture
positivepositive
negativenegative
identifyingidentifying
BlindBlind passagepassage22 - 3generatiogenerationn
negativenegative
Virus negatiVirus negativeve
asepsisasepsis
(一)(一) viral isolationviral isolation
11 animal inoculationanimal inoculation
chimpanzeemonkeyrabbitratchimpanzeemonkeyrabbitrat
22 embryonated egg inoculationembryonated egg inoculation
3 Cell culture3 Cell culture (( commonly usedcommonly used ))Primary and secondary cell culturePrimary and secondary cell culture
general proceduregeneral procedure
tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell
cell monolayercell monolayer
brokenbroken proteaseprotease
(primary cell)(primary cell)Serial subcultivationSerial subcultivation
Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)
features of virus cell culturefeatures of virus cell culture
A sensitive to many kinds of virusesA sensitive to many kinds of viruses
B cost highB cost high
C carry latent virusC carry latent virus
(2)Diploid(2)Diploid cellcell cultureculture
usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine
(3) secondary cell culture(3) secondary cell culture
sensitive to sensitive to multiplemultiple viruses viruses
high reproductive capacity long genera high reproductive capacity long generation timetion time
have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine
(( 二二 )) indexindex for reproduction of virusfor reproduction of virus
(1)Virus1)Virus infected cellinfected cell
(( cell roundinggathercell roundinggather
necrosisnecrosis ))
1 Cytopathic effect CPE)1 Cytopathic effect CPE)
CPECPE
(( 22 )) cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation
(( 33 )) viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell
CPECPE
22 hemadsorption hemadsorption HAd HAd
hemagglutininhemagglutinin (( HAHA )) on membrane of vion membrane of virus infected cell rus infected cell
Can Can adsorbadsorb vertebrate vertebrate RBCRBC
3 Interference3 Interference
4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange
( 三 ) viral quantitative assayHemagglutination test total amount
Plaque bacteriophage determination
ID50 or TCID50
Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le
sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex
cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a
nd hyphand hypha
22 cultureculture
specimen sabouraud mediumspecimen sabouraud medium
macroculture microculturemacroculture microculture
colonycolony
biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique
sporespore
3animal test3animal test
4skin test hypersensitivy4skin test hypersensitivy
(三)(三) mycology rapid diagnosismycology rapid diagnosis
serological test detect Ag or Abserological test detect Ag or Ab
detect nucleic acid of fungusdetect nucleic acid of fungus
detect mycotoxindetect mycotoxin
questionsquestions
11 definitiondefinition
serological identificationserological identification
and diagnosis)and diagnosis)
Cytopathic effect CPECytopathic effect CPE
hemadsorption HAdhemadsorption HAd
interferenceinterference
22 principle of Widalrsquos testprinciple of Widalrsquos test
33 principle of Gramrsquos stainingprinciple of Gramrsquos staining
Section 4Section 4
artificial active artificial active immunityimmunity
Artificial active immunityArtificial active immunity
stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid
vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA
A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow
th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni
citycity are repeatedly selected are repeatedly selected BCGBCG
The mutation is expressed only The mutation is expressed only
under certain conditionsunder certain conditions
may be may be useful in vaccinesuseful in vaccines
Conditional-lethal mutationsConditional-lethal mutations
temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation
the organisms can the organisms can replicate replicate at a at a
relativelyrelatively low low permissive temperature permissive temperature
but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive
temperaturetemperature
Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize
d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost
errdquo injectionerrdquo injection
A Live (attenuated) A Live (attenuated) vaccinesvaccines
B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate
responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe
toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves
artificial passive artificial passive immunization immunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
(( 22 )) staining techniquestaining technique
simple stainingsimple staining only use one dye only use one dye observe morphology arrangemen observe morphology arrangementt
compound stainingcompound staining use 2 kinds dyes or use 2 kinds dyes or more to identify bacteria more to identify bacteria
Gram stainingGram staining positivepositive (( violetviolet )) negativenegative (( redred ))Acid-fast stainingAcid-fast staining positivepositive (( redred )) negative negative (( blueblue ))Special stainingSpecial staining for bacterial capsule for bacterial capsule
spore flagella and so onspore flagella and so on
22 isolating culture and identifyisolating culture and identifyGeneral procedureGeneral procedure
specimen
Plain agar
Blood agar
Selective medium
cultural feature
nutrition
growing condition
Colony feature
hemolysis
pure
inoculate
stain initial identify (morphology)
Biochemical reaction
Serological identificationSerological identification species groupspecies groupss
Virulence Virulence identificationidentification
detect endotoxin or exotoxindetect endotoxin or exotoxin
animal testanimal test rabbit guinea pirabbit guinea pigg
Susceptibility Susceptibility testtest
Slip methodSlip method determine the naturedetermine the nature
Test tube methodTest tube method quantitationquantitation(( detect MBCdetect MBC MICMIC ))
33 detect Agdetect Ag
mdashmdash mdashmdashserological identificationserological identification
highly specificityhighly specificity
sensitivity sensitivity
44 other detectionother detection
Detect metabolic productDetect metabolic product
bull bull PCR detect DNAPCR detect DNA
caution avoid contaminationcaution avoid contamination (faulse positive)(faulse positive)
bull bull DNA chips techniqueDNA chips technique
22 serological diagnosisserological diagnosis
-----detect Ab with known Ag-----detect Ab with known Ag
------specimens------specimens serumserum
------paired serum (acute------paired serum (acute convalescent serum)convalescent serum)
Compared acute serum and convalescenCompared acute serum and convalescent serum quantitativelyt serum quantitatively
A 4-fold or greater increase in Ab titerA 4-fold or greater increase in Ab titer su supports a diagnosis of recent infectionpports a diagnosis of recent infection
Serum containing a high titer of Abs of thSerum containing a high titer of Abs of thee IgM IgM subclass would suggest a subclass would suggest a current icurrent infectionnfection
commonly used methodscommonly used methods
Agglutination testAgglutination test Widal test---diagnose typhus and Widal test---diagnose typhus and
paratyphoidparatyphoid
Well-Felix---diagnose rickettsiosisWell-Felix---diagnose rickettsiosis
10486801048680
Neutralization testNeutralization test antildquoOrdquo test---diagnose rheumatic antildquoOrdquo test---diagnose rheumatic
diseasedisease
Section 2 virology diagnosisSection 2 virology diagnosis
ViralViral infeinfectionction
Electron microscopeElectron microscope
Light microscope inLight microscope inclusion bodyclusion body
isolationisolation
VirusVirus particleparticle
V AgV Ag
V nucleic acidV nucleic acid
Viral enzymeViral enzyme retroviridaseretroviridase
DetectDetect virusvirus
DetectDetect AbAb
Choose the appropriate specimenChoose the appropriate specimen
Aseptic operationAseptic operation add antibiotics to speciadd antibiotics to speci
menmen
Take it in acute phase Take it in acute phase
Low temperature conservationLow temperature conservation transport it transport it
promptlypromptly preserve in 50preserve in 50 glycerineglycerine
Serologial diagnose paired serumSerologial diagnose paired serum
Specimen Selection Collection and ProcessingSpecimen Selection Collection and Processing
二二 viral isolation and identifyingviral isolation and identifying
specimenspecimenAnimal inoculaAnimal inoculationtion
Embryonated eEmbryonated egggg
Cell cultureCell culture
positivepositive
negativenegative
identifyingidentifying
BlindBlind passagepassage22 - 3generatiogenerationn
negativenegative
Virus negatiVirus negativeve
asepsisasepsis
(一)(一) viral isolationviral isolation
11 animal inoculationanimal inoculation
chimpanzeemonkeyrabbitratchimpanzeemonkeyrabbitrat
22 embryonated egg inoculationembryonated egg inoculation
3 Cell culture3 Cell culture (( commonly usedcommonly used ))Primary and secondary cell culturePrimary and secondary cell culture
general proceduregeneral procedure
tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell
cell monolayercell monolayer
brokenbroken proteaseprotease
(primary cell)(primary cell)Serial subcultivationSerial subcultivation
Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)
features of virus cell culturefeatures of virus cell culture
A sensitive to many kinds of virusesA sensitive to many kinds of viruses
B cost highB cost high
C carry latent virusC carry latent virus
(2)Diploid(2)Diploid cellcell cultureculture
usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine
(3) secondary cell culture(3) secondary cell culture
sensitive to sensitive to multiplemultiple viruses viruses
high reproductive capacity long genera high reproductive capacity long generation timetion time
have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine
(( 二二 )) indexindex for reproduction of virusfor reproduction of virus
(1)Virus1)Virus infected cellinfected cell
(( cell roundinggathercell roundinggather
necrosisnecrosis ))
1 Cytopathic effect CPE)1 Cytopathic effect CPE)
CPECPE
(( 22 )) cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation
(( 33 )) viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell
CPECPE
22 hemadsorption hemadsorption HAd HAd
hemagglutininhemagglutinin (( HAHA )) on membrane of vion membrane of virus infected cell rus infected cell
Can Can adsorbadsorb vertebrate vertebrate RBCRBC
3 Interference3 Interference
4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange
( 三 ) viral quantitative assayHemagglutination test total amount
Plaque bacteriophage determination
ID50 or TCID50
Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le
sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex
cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a
nd hyphand hypha
22 cultureculture
specimen sabouraud mediumspecimen sabouraud medium
macroculture microculturemacroculture microculture
colonycolony
biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique
sporespore
3animal test3animal test
4skin test hypersensitivy4skin test hypersensitivy
(三)(三) mycology rapid diagnosismycology rapid diagnosis
serological test detect Ag or Abserological test detect Ag or Ab
detect nucleic acid of fungusdetect nucleic acid of fungus
detect mycotoxindetect mycotoxin
questionsquestions
11 definitiondefinition
serological identificationserological identification
and diagnosis)and diagnosis)
Cytopathic effect CPECytopathic effect CPE
hemadsorption HAdhemadsorption HAd
interferenceinterference
22 principle of Widalrsquos testprinciple of Widalrsquos test
33 principle of Gramrsquos stainingprinciple of Gramrsquos staining
Section 4Section 4
artificial active artificial active immunityimmunity
Artificial active immunityArtificial active immunity
stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid
vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA
A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow
th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni
citycity are repeatedly selected are repeatedly selected BCGBCG
The mutation is expressed only The mutation is expressed only
under certain conditionsunder certain conditions
may be may be useful in vaccinesuseful in vaccines
Conditional-lethal mutationsConditional-lethal mutations
temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation
the organisms can the organisms can replicate replicate at a at a
relativelyrelatively low low permissive temperature permissive temperature
but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive
temperaturetemperature
Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize
d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost
errdquo injectionerrdquo injection
A Live (attenuated) A Live (attenuated) vaccinesvaccines
B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate
responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe
toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves
artificial passive artificial passive immunization immunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
Gram stainingGram staining positivepositive (( violetviolet )) negativenegative (( redred ))Acid-fast stainingAcid-fast staining positivepositive (( redred )) negative negative (( blueblue ))Special stainingSpecial staining for bacterial capsule for bacterial capsule
spore flagella and so onspore flagella and so on
22 isolating culture and identifyisolating culture and identifyGeneral procedureGeneral procedure
specimen
Plain agar
Blood agar
Selective medium
cultural feature
nutrition
growing condition
Colony feature
hemolysis
pure
inoculate
stain initial identify (morphology)
Biochemical reaction
Serological identificationSerological identification species groupspecies groupss
Virulence Virulence identificationidentification
detect endotoxin or exotoxindetect endotoxin or exotoxin
animal testanimal test rabbit guinea pirabbit guinea pigg
Susceptibility Susceptibility testtest
Slip methodSlip method determine the naturedetermine the nature
Test tube methodTest tube method quantitationquantitation(( detect MBCdetect MBC MICMIC ))
33 detect Agdetect Ag
mdashmdash mdashmdashserological identificationserological identification
highly specificityhighly specificity
sensitivity sensitivity
44 other detectionother detection
Detect metabolic productDetect metabolic product
bull bull PCR detect DNAPCR detect DNA
caution avoid contaminationcaution avoid contamination (faulse positive)(faulse positive)
bull bull DNA chips techniqueDNA chips technique
22 serological diagnosisserological diagnosis
-----detect Ab with known Ag-----detect Ab with known Ag
------specimens------specimens serumserum
------paired serum (acute------paired serum (acute convalescent serum)convalescent serum)
Compared acute serum and convalescenCompared acute serum and convalescent serum quantitativelyt serum quantitatively
A 4-fold or greater increase in Ab titerA 4-fold or greater increase in Ab titer su supports a diagnosis of recent infectionpports a diagnosis of recent infection
Serum containing a high titer of Abs of thSerum containing a high titer of Abs of thee IgM IgM subclass would suggest a subclass would suggest a current icurrent infectionnfection
commonly used methodscommonly used methods
Agglutination testAgglutination test Widal test---diagnose typhus and Widal test---diagnose typhus and
paratyphoidparatyphoid
Well-Felix---diagnose rickettsiosisWell-Felix---diagnose rickettsiosis
10486801048680
Neutralization testNeutralization test antildquoOrdquo test---diagnose rheumatic antildquoOrdquo test---diagnose rheumatic
diseasedisease
Section 2 virology diagnosisSection 2 virology diagnosis
ViralViral infeinfectionction
Electron microscopeElectron microscope
Light microscope inLight microscope inclusion bodyclusion body
isolationisolation
VirusVirus particleparticle
V AgV Ag
V nucleic acidV nucleic acid
Viral enzymeViral enzyme retroviridaseretroviridase
DetectDetect virusvirus
DetectDetect AbAb
Choose the appropriate specimenChoose the appropriate specimen
Aseptic operationAseptic operation add antibiotics to speciadd antibiotics to speci
menmen
Take it in acute phase Take it in acute phase
Low temperature conservationLow temperature conservation transport it transport it
promptlypromptly preserve in 50preserve in 50 glycerineglycerine
Serologial diagnose paired serumSerologial diagnose paired serum
Specimen Selection Collection and ProcessingSpecimen Selection Collection and Processing
二二 viral isolation and identifyingviral isolation and identifying
specimenspecimenAnimal inoculaAnimal inoculationtion
Embryonated eEmbryonated egggg
Cell cultureCell culture
positivepositive
negativenegative
identifyingidentifying
BlindBlind passagepassage22 - 3generatiogenerationn
negativenegative
Virus negatiVirus negativeve
asepsisasepsis
(一)(一) viral isolationviral isolation
11 animal inoculationanimal inoculation
chimpanzeemonkeyrabbitratchimpanzeemonkeyrabbitrat
22 embryonated egg inoculationembryonated egg inoculation
3 Cell culture3 Cell culture (( commonly usedcommonly used ))Primary and secondary cell culturePrimary and secondary cell culture
general proceduregeneral procedure
tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell
cell monolayercell monolayer
brokenbroken proteaseprotease
(primary cell)(primary cell)Serial subcultivationSerial subcultivation
Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)
features of virus cell culturefeatures of virus cell culture
A sensitive to many kinds of virusesA sensitive to many kinds of viruses
B cost highB cost high
C carry latent virusC carry latent virus
(2)Diploid(2)Diploid cellcell cultureculture
usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine
(3) secondary cell culture(3) secondary cell culture
sensitive to sensitive to multiplemultiple viruses viruses
high reproductive capacity long genera high reproductive capacity long generation timetion time
have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine
(( 二二 )) indexindex for reproduction of virusfor reproduction of virus
(1)Virus1)Virus infected cellinfected cell
(( cell roundinggathercell roundinggather
necrosisnecrosis ))
1 Cytopathic effect CPE)1 Cytopathic effect CPE)
CPECPE
(( 22 )) cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation
(( 33 )) viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell
CPECPE
22 hemadsorption hemadsorption HAd HAd
hemagglutininhemagglutinin (( HAHA )) on membrane of vion membrane of virus infected cell rus infected cell
Can Can adsorbadsorb vertebrate vertebrate RBCRBC
3 Interference3 Interference
4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange
( 三 ) viral quantitative assayHemagglutination test total amount
Plaque bacteriophage determination
ID50 or TCID50
Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le
sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex
cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a
nd hyphand hypha
22 cultureculture
specimen sabouraud mediumspecimen sabouraud medium
macroculture microculturemacroculture microculture
colonycolony
biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique
sporespore
3animal test3animal test
4skin test hypersensitivy4skin test hypersensitivy
(三)(三) mycology rapid diagnosismycology rapid diagnosis
serological test detect Ag or Abserological test detect Ag or Ab
detect nucleic acid of fungusdetect nucleic acid of fungus
detect mycotoxindetect mycotoxin
questionsquestions
11 definitiondefinition
serological identificationserological identification
and diagnosis)and diagnosis)
Cytopathic effect CPECytopathic effect CPE
hemadsorption HAdhemadsorption HAd
interferenceinterference
22 principle of Widalrsquos testprinciple of Widalrsquos test
33 principle of Gramrsquos stainingprinciple of Gramrsquos staining
Section 4Section 4
artificial active artificial active immunityimmunity
Artificial active immunityArtificial active immunity
stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid
vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA
A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow
th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni
citycity are repeatedly selected are repeatedly selected BCGBCG
The mutation is expressed only The mutation is expressed only
under certain conditionsunder certain conditions
may be may be useful in vaccinesuseful in vaccines
Conditional-lethal mutationsConditional-lethal mutations
temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation
the organisms can the organisms can replicate replicate at a at a
relativelyrelatively low low permissive temperature permissive temperature
but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive
temperaturetemperature
Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize
d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost
errdquo injectionerrdquo injection
A Live (attenuated) A Live (attenuated) vaccinesvaccines
B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate
responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe
toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves
artificial passive artificial passive immunization immunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
22 isolating culture and identifyisolating culture and identifyGeneral procedureGeneral procedure
specimen
Plain agar
Blood agar
Selective medium
cultural feature
nutrition
growing condition
Colony feature
hemolysis
pure
inoculate
stain initial identify (morphology)
Biochemical reaction
Serological identificationSerological identification species groupspecies groupss
Virulence Virulence identificationidentification
detect endotoxin or exotoxindetect endotoxin or exotoxin
animal testanimal test rabbit guinea pirabbit guinea pigg
Susceptibility Susceptibility testtest
Slip methodSlip method determine the naturedetermine the nature
Test tube methodTest tube method quantitationquantitation(( detect MBCdetect MBC MICMIC ))
33 detect Agdetect Ag
mdashmdash mdashmdashserological identificationserological identification
highly specificityhighly specificity
sensitivity sensitivity
44 other detectionother detection
Detect metabolic productDetect metabolic product
bull bull PCR detect DNAPCR detect DNA
caution avoid contaminationcaution avoid contamination (faulse positive)(faulse positive)
bull bull DNA chips techniqueDNA chips technique
22 serological diagnosisserological diagnosis
-----detect Ab with known Ag-----detect Ab with known Ag
------specimens------specimens serumserum
------paired serum (acute------paired serum (acute convalescent serum)convalescent serum)
Compared acute serum and convalescenCompared acute serum and convalescent serum quantitativelyt serum quantitatively
A 4-fold or greater increase in Ab titerA 4-fold or greater increase in Ab titer su supports a diagnosis of recent infectionpports a diagnosis of recent infection
Serum containing a high titer of Abs of thSerum containing a high titer of Abs of thee IgM IgM subclass would suggest a subclass would suggest a current icurrent infectionnfection
commonly used methodscommonly used methods
Agglutination testAgglutination test Widal test---diagnose typhus and Widal test---diagnose typhus and
paratyphoidparatyphoid
Well-Felix---diagnose rickettsiosisWell-Felix---diagnose rickettsiosis
10486801048680
Neutralization testNeutralization test antildquoOrdquo test---diagnose rheumatic antildquoOrdquo test---diagnose rheumatic
diseasedisease
Section 2 virology diagnosisSection 2 virology diagnosis
ViralViral infeinfectionction
Electron microscopeElectron microscope
Light microscope inLight microscope inclusion bodyclusion body
isolationisolation
VirusVirus particleparticle
V AgV Ag
V nucleic acidV nucleic acid
Viral enzymeViral enzyme retroviridaseretroviridase
DetectDetect virusvirus
DetectDetect AbAb
Choose the appropriate specimenChoose the appropriate specimen
Aseptic operationAseptic operation add antibiotics to speciadd antibiotics to speci
menmen
Take it in acute phase Take it in acute phase
Low temperature conservationLow temperature conservation transport it transport it
promptlypromptly preserve in 50preserve in 50 glycerineglycerine
Serologial diagnose paired serumSerologial diagnose paired serum
Specimen Selection Collection and ProcessingSpecimen Selection Collection and Processing
二二 viral isolation and identifyingviral isolation and identifying
specimenspecimenAnimal inoculaAnimal inoculationtion
Embryonated eEmbryonated egggg
Cell cultureCell culture
positivepositive
negativenegative
identifyingidentifying
BlindBlind passagepassage22 - 3generatiogenerationn
negativenegative
Virus negatiVirus negativeve
asepsisasepsis
(一)(一) viral isolationviral isolation
11 animal inoculationanimal inoculation
chimpanzeemonkeyrabbitratchimpanzeemonkeyrabbitrat
22 embryonated egg inoculationembryonated egg inoculation
3 Cell culture3 Cell culture (( commonly usedcommonly used ))Primary and secondary cell culturePrimary and secondary cell culture
general proceduregeneral procedure
tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell
cell monolayercell monolayer
brokenbroken proteaseprotease
(primary cell)(primary cell)Serial subcultivationSerial subcultivation
Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)
features of virus cell culturefeatures of virus cell culture
A sensitive to many kinds of virusesA sensitive to many kinds of viruses
B cost highB cost high
C carry latent virusC carry latent virus
(2)Diploid(2)Diploid cellcell cultureculture
usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine
(3) secondary cell culture(3) secondary cell culture
sensitive to sensitive to multiplemultiple viruses viruses
high reproductive capacity long genera high reproductive capacity long generation timetion time
have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine
(( 二二 )) indexindex for reproduction of virusfor reproduction of virus
(1)Virus1)Virus infected cellinfected cell
(( cell roundinggathercell roundinggather
necrosisnecrosis ))
1 Cytopathic effect CPE)1 Cytopathic effect CPE)
CPECPE
(( 22 )) cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation
(( 33 )) viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell
CPECPE
22 hemadsorption hemadsorption HAd HAd
hemagglutininhemagglutinin (( HAHA )) on membrane of vion membrane of virus infected cell rus infected cell
Can Can adsorbadsorb vertebrate vertebrate RBCRBC
3 Interference3 Interference
4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange
( 三 ) viral quantitative assayHemagglutination test total amount
Plaque bacteriophage determination
ID50 or TCID50
Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le
sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex
cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a
nd hyphand hypha
22 cultureculture
specimen sabouraud mediumspecimen sabouraud medium
macroculture microculturemacroculture microculture
colonycolony
biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique
sporespore
3animal test3animal test
4skin test hypersensitivy4skin test hypersensitivy
(三)(三) mycology rapid diagnosismycology rapid diagnosis
serological test detect Ag or Abserological test detect Ag or Ab
detect nucleic acid of fungusdetect nucleic acid of fungus
detect mycotoxindetect mycotoxin
questionsquestions
11 definitiondefinition
serological identificationserological identification
and diagnosis)and diagnosis)
Cytopathic effect CPECytopathic effect CPE
hemadsorption HAdhemadsorption HAd
interferenceinterference
22 principle of Widalrsquos testprinciple of Widalrsquos test
33 principle of Gramrsquos stainingprinciple of Gramrsquos staining
Section 4Section 4
artificial active artificial active immunityimmunity
Artificial active immunityArtificial active immunity
stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid
vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA
A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow
th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni
citycity are repeatedly selected are repeatedly selected BCGBCG
The mutation is expressed only The mutation is expressed only
under certain conditionsunder certain conditions
may be may be useful in vaccinesuseful in vaccines
Conditional-lethal mutationsConditional-lethal mutations
temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation
the organisms can the organisms can replicate replicate at a at a
relativelyrelatively low low permissive temperature permissive temperature
but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive
temperaturetemperature
Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize
d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost
errdquo injectionerrdquo injection
A Live (attenuated) A Live (attenuated) vaccinesvaccines
B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate
responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe
toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves
artificial passive artificial passive immunization immunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
Serological identificationSerological identification species groupspecies groupss
Virulence Virulence identificationidentification
detect endotoxin or exotoxindetect endotoxin or exotoxin
animal testanimal test rabbit guinea pirabbit guinea pigg
Susceptibility Susceptibility testtest
Slip methodSlip method determine the naturedetermine the nature
Test tube methodTest tube method quantitationquantitation(( detect MBCdetect MBC MICMIC ))
33 detect Agdetect Ag
mdashmdash mdashmdashserological identificationserological identification
highly specificityhighly specificity
sensitivity sensitivity
44 other detectionother detection
Detect metabolic productDetect metabolic product
bull bull PCR detect DNAPCR detect DNA
caution avoid contaminationcaution avoid contamination (faulse positive)(faulse positive)
bull bull DNA chips techniqueDNA chips technique
22 serological diagnosisserological diagnosis
-----detect Ab with known Ag-----detect Ab with known Ag
------specimens------specimens serumserum
------paired serum (acute------paired serum (acute convalescent serum)convalescent serum)
Compared acute serum and convalescenCompared acute serum and convalescent serum quantitativelyt serum quantitatively
A 4-fold or greater increase in Ab titerA 4-fold or greater increase in Ab titer su supports a diagnosis of recent infectionpports a diagnosis of recent infection
Serum containing a high titer of Abs of thSerum containing a high titer of Abs of thee IgM IgM subclass would suggest a subclass would suggest a current icurrent infectionnfection
commonly used methodscommonly used methods
Agglutination testAgglutination test Widal test---diagnose typhus and Widal test---diagnose typhus and
paratyphoidparatyphoid
Well-Felix---diagnose rickettsiosisWell-Felix---diagnose rickettsiosis
10486801048680
Neutralization testNeutralization test antildquoOrdquo test---diagnose rheumatic antildquoOrdquo test---diagnose rheumatic
diseasedisease
Section 2 virology diagnosisSection 2 virology diagnosis
ViralViral infeinfectionction
Electron microscopeElectron microscope
Light microscope inLight microscope inclusion bodyclusion body
isolationisolation
VirusVirus particleparticle
V AgV Ag
V nucleic acidV nucleic acid
Viral enzymeViral enzyme retroviridaseretroviridase
DetectDetect virusvirus
DetectDetect AbAb
Choose the appropriate specimenChoose the appropriate specimen
Aseptic operationAseptic operation add antibiotics to speciadd antibiotics to speci
menmen
Take it in acute phase Take it in acute phase
Low temperature conservationLow temperature conservation transport it transport it
promptlypromptly preserve in 50preserve in 50 glycerineglycerine
Serologial diagnose paired serumSerologial diagnose paired serum
Specimen Selection Collection and ProcessingSpecimen Selection Collection and Processing
二二 viral isolation and identifyingviral isolation and identifying
specimenspecimenAnimal inoculaAnimal inoculationtion
Embryonated eEmbryonated egggg
Cell cultureCell culture
positivepositive
negativenegative
identifyingidentifying
BlindBlind passagepassage22 - 3generatiogenerationn
negativenegative
Virus negatiVirus negativeve
asepsisasepsis
(一)(一) viral isolationviral isolation
11 animal inoculationanimal inoculation
chimpanzeemonkeyrabbitratchimpanzeemonkeyrabbitrat
22 embryonated egg inoculationembryonated egg inoculation
3 Cell culture3 Cell culture (( commonly usedcommonly used ))Primary and secondary cell culturePrimary and secondary cell culture
general proceduregeneral procedure
tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell
cell monolayercell monolayer
brokenbroken proteaseprotease
(primary cell)(primary cell)Serial subcultivationSerial subcultivation
Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)
features of virus cell culturefeatures of virus cell culture
A sensitive to many kinds of virusesA sensitive to many kinds of viruses
B cost highB cost high
C carry latent virusC carry latent virus
(2)Diploid(2)Diploid cellcell cultureculture
usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine
(3) secondary cell culture(3) secondary cell culture
sensitive to sensitive to multiplemultiple viruses viruses
high reproductive capacity long genera high reproductive capacity long generation timetion time
have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine
(( 二二 )) indexindex for reproduction of virusfor reproduction of virus
(1)Virus1)Virus infected cellinfected cell
(( cell roundinggathercell roundinggather
necrosisnecrosis ))
1 Cytopathic effect CPE)1 Cytopathic effect CPE)
CPECPE
(( 22 )) cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation
(( 33 )) viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell
CPECPE
22 hemadsorption hemadsorption HAd HAd
hemagglutininhemagglutinin (( HAHA )) on membrane of vion membrane of virus infected cell rus infected cell
Can Can adsorbadsorb vertebrate vertebrate RBCRBC
3 Interference3 Interference
4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange
( 三 ) viral quantitative assayHemagglutination test total amount
Plaque bacteriophage determination
ID50 or TCID50
Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le
sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex
cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a
nd hyphand hypha
22 cultureculture
specimen sabouraud mediumspecimen sabouraud medium
macroculture microculturemacroculture microculture
colonycolony
biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique
sporespore
3animal test3animal test
4skin test hypersensitivy4skin test hypersensitivy
(三)(三) mycology rapid diagnosismycology rapid diagnosis
serological test detect Ag or Abserological test detect Ag or Ab
detect nucleic acid of fungusdetect nucleic acid of fungus
detect mycotoxindetect mycotoxin
questionsquestions
11 definitiondefinition
serological identificationserological identification
and diagnosis)and diagnosis)
Cytopathic effect CPECytopathic effect CPE
hemadsorption HAdhemadsorption HAd
interferenceinterference
22 principle of Widalrsquos testprinciple of Widalrsquos test
33 principle of Gramrsquos stainingprinciple of Gramrsquos staining
Section 4Section 4
artificial active artificial active immunityimmunity
Artificial active immunityArtificial active immunity
stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid
vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA
A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow
th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni
citycity are repeatedly selected are repeatedly selected BCGBCG
The mutation is expressed only The mutation is expressed only
under certain conditionsunder certain conditions
may be may be useful in vaccinesuseful in vaccines
Conditional-lethal mutationsConditional-lethal mutations
temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation
the organisms can the organisms can replicate replicate at a at a
relativelyrelatively low low permissive temperature permissive temperature
but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive
temperaturetemperature
Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize
d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost
errdquo injectionerrdquo injection
A Live (attenuated) A Live (attenuated) vaccinesvaccines
B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate
responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe
toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves
artificial passive artificial passive immunization immunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
33 detect Agdetect Ag
mdashmdash mdashmdashserological identificationserological identification
highly specificityhighly specificity
sensitivity sensitivity
44 other detectionother detection
Detect metabolic productDetect metabolic product
bull bull PCR detect DNAPCR detect DNA
caution avoid contaminationcaution avoid contamination (faulse positive)(faulse positive)
bull bull DNA chips techniqueDNA chips technique
22 serological diagnosisserological diagnosis
-----detect Ab with known Ag-----detect Ab with known Ag
------specimens------specimens serumserum
------paired serum (acute------paired serum (acute convalescent serum)convalescent serum)
Compared acute serum and convalescenCompared acute serum and convalescent serum quantitativelyt serum quantitatively
A 4-fold or greater increase in Ab titerA 4-fold or greater increase in Ab titer su supports a diagnosis of recent infectionpports a diagnosis of recent infection
Serum containing a high titer of Abs of thSerum containing a high titer of Abs of thee IgM IgM subclass would suggest a subclass would suggest a current icurrent infectionnfection
commonly used methodscommonly used methods
Agglutination testAgglutination test Widal test---diagnose typhus and Widal test---diagnose typhus and
paratyphoidparatyphoid
Well-Felix---diagnose rickettsiosisWell-Felix---diagnose rickettsiosis
10486801048680
Neutralization testNeutralization test antildquoOrdquo test---diagnose rheumatic antildquoOrdquo test---diagnose rheumatic
diseasedisease
Section 2 virology diagnosisSection 2 virology diagnosis
ViralViral infeinfectionction
Electron microscopeElectron microscope
Light microscope inLight microscope inclusion bodyclusion body
isolationisolation
VirusVirus particleparticle
V AgV Ag
V nucleic acidV nucleic acid
Viral enzymeViral enzyme retroviridaseretroviridase
DetectDetect virusvirus
DetectDetect AbAb
Choose the appropriate specimenChoose the appropriate specimen
Aseptic operationAseptic operation add antibiotics to speciadd antibiotics to speci
menmen
Take it in acute phase Take it in acute phase
Low temperature conservationLow temperature conservation transport it transport it
promptlypromptly preserve in 50preserve in 50 glycerineglycerine
Serologial diagnose paired serumSerologial diagnose paired serum
Specimen Selection Collection and ProcessingSpecimen Selection Collection and Processing
二二 viral isolation and identifyingviral isolation and identifying
specimenspecimenAnimal inoculaAnimal inoculationtion
Embryonated eEmbryonated egggg
Cell cultureCell culture
positivepositive
negativenegative
identifyingidentifying
BlindBlind passagepassage22 - 3generatiogenerationn
negativenegative
Virus negatiVirus negativeve
asepsisasepsis
(一)(一) viral isolationviral isolation
11 animal inoculationanimal inoculation
chimpanzeemonkeyrabbitratchimpanzeemonkeyrabbitrat
22 embryonated egg inoculationembryonated egg inoculation
3 Cell culture3 Cell culture (( commonly usedcommonly used ))Primary and secondary cell culturePrimary and secondary cell culture
general proceduregeneral procedure
tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell
cell monolayercell monolayer
brokenbroken proteaseprotease
(primary cell)(primary cell)Serial subcultivationSerial subcultivation
Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)
features of virus cell culturefeatures of virus cell culture
A sensitive to many kinds of virusesA sensitive to many kinds of viruses
B cost highB cost high
C carry latent virusC carry latent virus
(2)Diploid(2)Diploid cellcell cultureculture
usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine
(3) secondary cell culture(3) secondary cell culture
sensitive to sensitive to multiplemultiple viruses viruses
high reproductive capacity long genera high reproductive capacity long generation timetion time
have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine
(( 二二 )) indexindex for reproduction of virusfor reproduction of virus
(1)Virus1)Virus infected cellinfected cell
(( cell roundinggathercell roundinggather
necrosisnecrosis ))
1 Cytopathic effect CPE)1 Cytopathic effect CPE)
CPECPE
(( 22 )) cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation
(( 33 )) viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell
CPECPE
22 hemadsorption hemadsorption HAd HAd
hemagglutininhemagglutinin (( HAHA )) on membrane of vion membrane of virus infected cell rus infected cell
Can Can adsorbadsorb vertebrate vertebrate RBCRBC
3 Interference3 Interference
4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange
( 三 ) viral quantitative assayHemagglutination test total amount
Plaque bacteriophage determination
ID50 or TCID50
Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le
sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex
cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a
nd hyphand hypha
22 cultureculture
specimen sabouraud mediumspecimen sabouraud medium
macroculture microculturemacroculture microculture
colonycolony
biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique
sporespore
3animal test3animal test
4skin test hypersensitivy4skin test hypersensitivy
(三)(三) mycology rapid diagnosismycology rapid diagnosis
serological test detect Ag or Abserological test detect Ag or Ab
detect nucleic acid of fungusdetect nucleic acid of fungus
detect mycotoxindetect mycotoxin
questionsquestions
11 definitiondefinition
serological identificationserological identification
and diagnosis)and diagnosis)
Cytopathic effect CPECytopathic effect CPE
hemadsorption HAdhemadsorption HAd
interferenceinterference
22 principle of Widalrsquos testprinciple of Widalrsquos test
33 principle of Gramrsquos stainingprinciple of Gramrsquos staining
Section 4Section 4
artificial active artificial active immunityimmunity
Artificial active immunityArtificial active immunity
stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid
vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA
A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow
th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni
citycity are repeatedly selected are repeatedly selected BCGBCG
The mutation is expressed only The mutation is expressed only
under certain conditionsunder certain conditions
may be may be useful in vaccinesuseful in vaccines
Conditional-lethal mutationsConditional-lethal mutations
temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation
the organisms can the organisms can replicate replicate at a at a
relativelyrelatively low low permissive temperature permissive temperature
but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive
temperaturetemperature
Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize
d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost
errdquo injectionerrdquo injection
A Live (attenuated) A Live (attenuated) vaccinesvaccines
B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate
responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe
toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves
artificial passive artificial passive immunization immunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
44 other detectionother detection
Detect metabolic productDetect metabolic product
bull bull PCR detect DNAPCR detect DNA
caution avoid contaminationcaution avoid contamination (faulse positive)(faulse positive)
bull bull DNA chips techniqueDNA chips technique
22 serological diagnosisserological diagnosis
-----detect Ab with known Ag-----detect Ab with known Ag
------specimens------specimens serumserum
------paired serum (acute------paired serum (acute convalescent serum)convalescent serum)
Compared acute serum and convalescenCompared acute serum and convalescent serum quantitativelyt serum quantitatively
A 4-fold or greater increase in Ab titerA 4-fold or greater increase in Ab titer su supports a diagnosis of recent infectionpports a diagnosis of recent infection
Serum containing a high titer of Abs of thSerum containing a high titer of Abs of thee IgM IgM subclass would suggest a subclass would suggest a current icurrent infectionnfection
commonly used methodscommonly used methods
Agglutination testAgglutination test Widal test---diagnose typhus and Widal test---diagnose typhus and
paratyphoidparatyphoid
Well-Felix---diagnose rickettsiosisWell-Felix---diagnose rickettsiosis
10486801048680
Neutralization testNeutralization test antildquoOrdquo test---diagnose rheumatic antildquoOrdquo test---diagnose rheumatic
diseasedisease
Section 2 virology diagnosisSection 2 virology diagnosis
ViralViral infeinfectionction
Electron microscopeElectron microscope
Light microscope inLight microscope inclusion bodyclusion body
isolationisolation
VirusVirus particleparticle
V AgV Ag
V nucleic acidV nucleic acid
Viral enzymeViral enzyme retroviridaseretroviridase
DetectDetect virusvirus
DetectDetect AbAb
Choose the appropriate specimenChoose the appropriate specimen
Aseptic operationAseptic operation add antibiotics to speciadd antibiotics to speci
menmen
Take it in acute phase Take it in acute phase
Low temperature conservationLow temperature conservation transport it transport it
promptlypromptly preserve in 50preserve in 50 glycerineglycerine
Serologial diagnose paired serumSerologial diagnose paired serum
Specimen Selection Collection and ProcessingSpecimen Selection Collection and Processing
二二 viral isolation and identifyingviral isolation and identifying
specimenspecimenAnimal inoculaAnimal inoculationtion
Embryonated eEmbryonated egggg
Cell cultureCell culture
positivepositive
negativenegative
identifyingidentifying
BlindBlind passagepassage22 - 3generatiogenerationn
negativenegative
Virus negatiVirus negativeve
asepsisasepsis
(一)(一) viral isolationviral isolation
11 animal inoculationanimal inoculation
chimpanzeemonkeyrabbitratchimpanzeemonkeyrabbitrat
22 embryonated egg inoculationembryonated egg inoculation
3 Cell culture3 Cell culture (( commonly usedcommonly used ))Primary and secondary cell culturePrimary and secondary cell culture
general proceduregeneral procedure
tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell
cell monolayercell monolayer
brokenbroken proteaseprotease
(primary cell)(primary cell)Serial subcultivationSerial subcultivation
Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)
features of virus cell culturefeatures of virus cell culture
A sensitive to many kinds of virusesA sensitive to many kinds of viruses
B cost highB cost high
C carry latent virusC carry latent virus
(2)Diploid(2)Diploid cellcell cultureculture
usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine
(3) secondary cell culture(3) secondary cell culture
sensitive to sensitive to multiplemultiple viruses viruses
high reproductive capacity long genera high reproductive capacity long generation timetion time
have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine
(( 二二 )) indexindex for reproduction of virusfor reproduction of virus
(1)Virus1)Virus infected cellinfected cell
(( cell roundinggathercell roundinggather
necrosisnecrosis ))
1 Cytopathic effect CPE)1 Cytopathic effect CPE)
CPECPE
(( 22 )) cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation
(( 33 )) viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell
CPECPE
22 hemadsorption hemadsorption HAd HAd
hemagglutininhemagglutinin (( HAHA )) on membrane of vion membrane of virus infected cell rus infected cell
Can Can adsorbadsorb vertebrate vertebrate RBCRBC
3 Interference3 Interference
4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange
( 三 ) viral quantitative assayHemagglutination test total amount
Plaque bacteriophage determination
ID50 or TCID50
Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le
sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex
cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a
nd hyphand hypha
22 cultureculture
specimen sabouraud mediumspecimen sabouraud medium
macroculture microculturemacroculture microculture
colonycolony
biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique
sporespore
3animal test3animal test
4skin test hypersensitivy4skin test hypersensitivy
(三)(三) mycology rapid diagnosismycology rapid diagnosis
serological test detect Ag or Abserological test detect Ag or Ab
detect nucleic acid of fungusdetect nucleic acid of fungus
detect mycotoxindetect mycotoxin
questionsquestions
11 definitiondefinition
serological identificationserological identification
and diagnosis)and diagnosis)
Cytopathic effect CPECytopathic effect CPE
hemadsorption HAdhemadsorption HAd
interferenceinterference
22 principle of Widalrsquos testprinciple of Widalrsquos test
33 principle of Gramrsquos stainingprinciple of Gramrsquos staining
Section 4Section 4
artificial active artificial active immunityimmunity
Artificial active immunityArtificial active immunity
stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid
vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA
A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow
th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni
citycity are repeatedly selected are repeatedly selected BCGBCG
The mutation is expressed only The mutation is expressed only
under certain conditionsunder certain conditions
may be may be useful in vaccinesuseful in vaccines
Conditional-lethal mutationsConditional-lethal mutations
temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation
the organisms can the organisms can replicate replicate at a at a
relativelyrelatively low low permissive temperature permissive temperature
but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive
temperaturetemperature
Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize
d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost
errdquo injectionerrdquo injection
A Live (attenuated) A Live (attenuated) vaccinesvaccines
B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate
responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe
toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves
artificial passive artificial passive immunization immunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
22 serological diagnosisserological diagnosis
-----detect Ab with known Ag-----detect Ab with known Ag
------specimens------specimens serumserum
------paired serum (acute------paired serum (acute convalescent serum)convalescent serum)
Compared acute serum and convalescenCompared acute serum and convalescent serum quantitativelyt serum quantitatively
A 4-fold or greater increase in Ab titerA 4-fold or greater increase in Ab titer su supports a diagnosis of recent infectionpports a diagnosis of recent infection
Serum containing a high titer of Abs of thSerum containing a high titer of Abs of thee IgM IgM subclass would suggest a subclass would suggest a current icurrent infectionnfection
commonly used methodscommonly used methods
Agglutination testAgglutination test Widal test---diagnose typhus and Widal test---diagnose typhus and
paratyphoidparatyphoid
Well-Felix---diagnose rickettsiosisWell-Felix---diagnose rickettsiosis
10486801048680
Neutralization testNeutralization test antildquoOrdquo test---diagnose rheumatic antildquoOrdquo test---diagnose rheumatic
diseasedisease
Section 2 virology diagnosisSection 2 virology diagnosis
ViralViral infeinfectionction
Electron microscopeElectron microscope
Light microscope inLight microscope inclusion bodyclusion body
isolationisolation
VirusVirus particleparticle
V AgV Ag
V nucleic acidV nucleic acid
Viral enzymeViral enzyme retroviridaseretroviridase
DetectDetect virusvirus
DetectDetect AbAb
Choose the appropriate specimenChoose the appropriate specimen
Aseptic operationAseptic operation add antibiotics to speciadd antibiotics to speci
menmen
Take it in acute phase Take it in acute phase
Low temperature conservationLow temperature conservation transport it transport it
promptlypromptly preserve in 50preserve in 50 glycerineglycerine
Serologial diagnose paired serumSerologial diagnose paired serum
Specimen Selection Collection and ProcessingSpecimen Selection Collection and Processing
二二 viral isolation and identifyingviral isolation and identifying
specimenspecimenAnimal inoculaAnimal inoculationtion
Embryonated eEmbryonated egggg
Cell cultureCell culture
positivepositive
negativenegative
identifyingidentifying
BlindBlind passagepassage22 - 3generatiogenerationn
negativenegative
Virus negatiVirus negativeve
asepsisasepsis
(一)(一) viral isolationviral isolation
11 animal inoculationanimal inoculation
chimpanzeemonkeyrabbitratchimpanzeemonkeyrabbitrat
22 embryonated egg inoculationembryonated egg inoculation
3 Cell culture3 Cell culture (( commonly usedcommonly used ))Primary and secondary cell culturePrimary and secondary cell culture
general proceduregeneral procedure
tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell
cell monolayercell monolayer
brokenbroken proteaseprotease
(primary cell)(primary cell)Serial subcultivationSerial subcultivation
Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)
features of virus cell culturefeatures of virus cell culture
A sensitive to many kinds of virusesA sensitive to many kinds of viruses
B cost highB cost high
C carry latent virusC carry latent virus
(2)Diploid(2)Diploid cellcell cultureculture
usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine
(3) secondary cell culture(3) secondary cell culture
sensitive to sensitive to multiplemultiple viruses viruses
high reproductive capacity long genera high reproductive capacity long generation timetion time
have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine
(( 二二 )) indexindex for reproduction of virusfor reproduction of virus
(1)Virus1)Virus infected cellinfected cell
(( cell roundinggathercell roundinggather
necrosisnecrosis ))
1 Cytopathic effect CPE)1 Cytopathic effect CPE)
CPECPE
(( 22 )) cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation
(( 33 )) viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell
CPECPE
22 hemadsorption hemadsorption HAd HAd
hemagglutininhemagglutinin (( HAHA )) on membrane of vion membrane of virus infected cell rus infected cell
Can Can adsorbadsorb vertebrate vertebrate RBCRBC
3 Interference3 Interference
4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange
( 三 ) viral quantitative assayHemagglutination test total amount
Plaque bacteriophage determination
ID50 or TCID50
Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le
sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex
cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a
nd hyphand hypha
22 cultureculture
specimen sabouraud mediumspecimen sabouraud medium
macroculture microculturemacroculture microculture
colonycolony
biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique
sporespore
3animal test3animal test
4skin test hypersensitivy4skin test hypersensitivy
(三)(三) mycology rapid diagnosismycology rapid diagnosis
serological test detect Ag or Abserological test detect Ag or Ab
detect nucleic acid of fungusdetect nucleic acid of fungus
detect mycotoxindetect mycotoxin
questionsquestions
11 definitiondefinition
serological identificationserological identification
and diagnosis)and diagnosis)
Cytopathic effect CPECytopathic effect CPE
hemadsorption HAdhemadsorption HAd
interferenceinterference
22 principle of Widalrsquos testprinciple of Widalrsquos test
33 principle of Gramrsquos stainingprinciple of Gramrsquos staining
Section 4Section 4
artificial active artificial active immunityimmunity
Artificial active immunityArtificial active immunity
stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid
vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA
A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow
th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni
citycity are repeatedly selected are repeatedly selected BCGBCG
The mutation is expressed only The mutation is expressed only
under certain conditionsunder certain conditions
may be may be useful in vaccinesuseful in vaccines
Conditional-lethal mutationsConditional-lethal mutations
temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation
the organisms can the organisms can replicate replicate at a at a
relativelyrelatively low low permissive temperature permissive temperature
but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive
temperaturetemperature
Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize
d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost
errdquo injectionerrdquo injection
A Live (attenuated) A Live (attenuated) vaccinesvaccines
B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate
responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe
toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves
artificial passive artificial passive immunization immunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
Compared acute serum and convalescenCompared acute serum and convalescent serum quantitativelyt serum quantitatively
A 4-fold or greater increase in Ab titerA 4-fold or greater increase in Ab titer su supports a diagnosis of recent infectionpports a diagnosis of recent infection
Serum containing a high titer of Abs of thSerum containing a high titer of Abs of thee IgM IgM subclass would suggest a subclass would suggest a current icurrent infectionnfection
commonly used methodscommonly used methods
Agglutination testAgglutination test Widal test---diagnose typhus and Widal test---diagnose typhus and
paratyphoidparatyphoid
Well-Felix---diagnose rickettsiosisWell-Felix---diagnose rickettsiosis
10486801048680
Neutralization testNeutralization test antildquoOrdquo test---diagnose rheumatic antildquoOrdquo test---diagnose rheumatic
diseasedisease
Section 2 virology diagnosisSection 2 virology diagnosis
ViralViral infeinfectionction
Electron microscopeElectron microscope
Light microscope inLight microscope inclusion bodyclusion body
isolationisolation
VirusVirus particleparticle
V AgV Ag
V nucleic acidV nucleic acid
Viral enzymeViral enzyme retroviridaseretroviridase
DetectDetect virusvirus
DetectDetect AbAb
Choose the appropriate specimenChoose the appropriate specimen
Aseptic operationAseptic operation add antibiotics to speciadd antibiotics to speci
menmen
Take it in acute phase Take it in acute phase
Low temperature conservationLow temperature conservation transport it transport it
promptlypromptly preserve in 50preserve in 50 glycerineglycerine
Serologial diagnose paired serumSerologial diagnose paired serum
Specimen Selection Collection and ProcessingSpecimen Selection Collection and Processing
二二 viral isolation and identifyingviral isolation and identifying
specimenspecimenAnimal inoculaAnimal inoculationtion
Embryonated eEmbryonated egggg
Cell cultureCell culture
positivepositive
negativenegative
identifyingidentifying
BlindBlind passagepassage22 - 3generatiogenerationn
negativenegative
Virus negatiVirus negativeve
asepsisasepsis
(一)(一) viral isolationviral isolation
11 animal inoculationanimal inoculation
chimpanzeemonkeyrabbitratchimpanzeemonkeyrabbitrat
22 embryonated egg inoculationembryonated egg inoculation
3 Cell culture3 Cell culture (( commonly usedcommonly used ))Primary and secondary cell culturePrimary and secondary cell culture
general proceduregeneral procedure
tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell
cell monolayercell monolayer
brokenbroken proteaseprotease
(primary cell)(primary cell)Serial subcultivationSerial subcultivation
Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)
features of virus cell culturefeatures of virus cell culture
A sensitive to many kinds of virusesA sensitive to many kinds of viruses
B cost highB cost high
C carry latent virusC carry latent virus
(2)Diploid(2)Diploid cellcell cultureculture
usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine
(3) secondary cell culture(3) secondary cell culture
sensitive to sensitive to multiplemultiple viruses viruses
high reproductive capacity long genera high reproductive capacity long generation timetion time
have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine
(( 二二 )) indexindex for reproduction of virusfor reproduction of virus
(1)Virus1)Virus infected cellinfected cell
(( cell roundinggathercell roundinggather
necrosisnecrosis ))
1 Cytopathic effect CPE)1 Cytopathic effect CPE)
CPECPE
(( 22 )) cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation
(( 33 )) viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell
CPECPE
22 hemadsorption hemadsorption HAd HAd
hemagglutininhemagglutinin (( HAHA )) on membrane of vion membrane of virus infected cell rus infected cell
Can Can adsorbadsorb vertebrate vertebrate RBCRBC
3 Interference3 Interference
4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange
( 三 ) viral quantitative assayHemagglutination test total amount
Plaque bacteriophage determination
ID50 or TCID50
Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le
sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex
cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a
nd hyphand hypha
22 cultureculture
specimen sabouraud mediumspecimen sabouraud medium
macroculture microculturemacroculture microculture
colonycolony
biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique
sporespore
3animal test3animal test
4skin test hypersensitivy4skin test hypersensitivy
(三)(三) mycology rapid diagnosismycology rapid diagnosis
serological test detect Ag or Abserological test detect Ag or Ab
detect nucleic acid of fungusdetect nucleic acid of fungus
detect mycotoxindetect mycotoxin
questionsquestions
11 definitiondefinition
serological identificationserological identification
and diagnosis)and diagnosis)
Cytopathic effect CPECytopathic effect CPE
hemadsorption HAdhemadsorption HAd
interferenceinterference
22 principle of Widalrsquos testprinciple of Widalrsquos test
33 principle of Gramrsquos stainingprinciple of Gramrsquos staining
Section 4Section 4
artificial active artificial active immunityimmunity
Artificial active immunityArtificial active immunity
stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid
vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA
A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow
th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni
citycity are repeatedly selected are repeatedly selected BCGBCG
The mutation is expressed only The mutation is expressed only
under certain conditionsunder certain conditions
may be may be useful in vaccinesuseful in vaccines
Conditional-lethal mutationsConditional-lethal mutations
temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation
the organisms can the organisms can replicate replicate at a at a
relativelyrelatively low low permissive temperature permissive temperature
but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive
temperaturetemperature
Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize
d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost
errdquo injectionerrdquo injection
A Live (attenuated) A Live (attenuated) vaccinesvaccines
B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate
responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe
toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves
artificial passive artificial passive immunization immunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
commonly used methodscommonly used methods
Agglutination testAgglutination test Widal test---diagnose typhus and Widal test---diagnose typhus and
paratyphoidparatyphoid
Well-Felix---diagnose rickettsiosisWell-Felix---diagnose rickettsiosis
10486801048680
Neutralization testNeutralization test antildquoOrdquo test---diagnose rheumatic antildquoOrdquo test---diagnose rheumatic
diseasedisease
Section 2 virology diagnosisSection 2 virology diagnosis
ViralViral infeinfectionction
Electron microscopeElectron microscope
Light microscope inLight microscope inclusion bodyclusion body
isolationisolation
VirusVirus particleparticle
V AgV Ag
V nucleic acidV nucleic acid
Viral enzymeViral enzyme retroviridaseretroviridase
DetectDetect virusvirus
DetectDetect AbAb
Choose the appropriate specimenChoose the appropriate specimen
Aseptic operationAseptic operation add antibiotics to speciadd antibiotics to speci
menmen
Take it in acute phase Take it in acute phase
Low temperature conservationLow temperature conservation transport it transport it
promptlypromptly preserve in 50preserve in 50 glycerineglycerine
Serologial diagnose paired serumSerologial diagnose paired serum
Specimen Selection Collection and ProcessingSpecimen Selection Collection and Processing
二二 viral isolation and identifyingviral isolation and identifying
specimenspecimenAnimal inoculaAnimal inoculationtion
Embryonated eEmbryonated egggg
Cell cultureCell culture
positivepositive
negativenegative
identifyingidentifying
BlindBlind passagepassage22 - 3generatiogenerationn
negativenegative
Virus negatiVirus negativeve
asepsisasepsis
(一)(一) viral isolationviral isolation
11 animal inoculationanimal inoculation
chimpanzeemonkeyrabbitratchimpanzeemonkeyrabbitrat
22 embryonated egg inoculationembryonated egg inoculation
3 Cell culture3 Cell culture (( commonly usedcommonly used ))Primary and secondary cell culturePrimary and secondary cell culture
general proceduregeneral procedure
tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell
cell monolayercell monolayer
brokenbroken proteaseprotease
(primary cell)(primary cell)Serial subcultivationSerial subcultivation
Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)
features of virus cell culturefeatures of virus cell culture
A sensitive to many kinds of virusesA sensitive to many kinds of viruses
B cost highB cost high
C carry latent virusC carry latent virus
(2)Diploid(2)Diploid cellcell cultureculture
usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine
(3) secondary cell culture(3) secondary cell culture
sensitive to sensitive to multiplemultiple viruses viruses
high reproductive capacity long genera high reproductive capacity long generation timetion time
have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine
(( 二二 )) indexindex for reproduction of virusfor reproduction of virus
(1)Virus1)Virus infected cellinfected cell
(( cell roundinggathercell roundinggather
necrosisnecrosis ))
1 Cytopathic effect CPE)1 Cytopathic effect CPE)
CPECPE
(( 22 )) cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation
(( 33 )) viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell
CPECPE
22 hemadsorption hemadsorption HAd HAd
hemagglutininhemagglutinin (( HAHA )) on membrane of vion membrane of virus infected cell rus infected cell
Can Can adsorbadsorb vertebrate vertebrate RBCRBC
3 Interference3 Interference
4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange
( 三 ) viral quantitative assayHemagglutination test total amount
Plaque bacteriophage determination
ID50 or TCID50
Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le
sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex
cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a
nd hyphand hypha
22 cultureculture
specimen sabouraud mediumspecimen sabouraud medium
macroculture microculturemacroculture microculture
colonycolony
biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique
sporespore
3animal test3animal test
4skin test hypersensitivy4skin test hypersensitivy
(三)(三) mycology rapid diagnosismycology rapid diagnosis
serological test detect Ag or Abserological test detect Ag or Ab
detect nucleic acid of fungusdetect nucleic acid of fungus
detect mycotoxindetect mycotoxin
questionsquestions
11 definitiondefinition
serological identificationserological identification
and diagnosis)and diagnosis)
Cytopathic effect CPECytopathic effect CPE
hemadsorption HAdhemadsorption HAd
interferenceinterference
22 principle of Widalrsquos testprinciple of Widalrsquos test
33 principle of Gramrsquos stainingprinciple of Gramrsquos staining
Section 4Section 4
artificial active artificial active immunityimmunity
Artificial active immunityArtificial active immunity
stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid
vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA
A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow
th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni
citycity are repeatedly selected are repeatedly selected BCGBCG
The mutation is expressed only The mutation is expressed only
under certain conditionsunder certain conditions
may be may be useful in vaccinesuseful in vaccines
Conditional-lethal mutationsConditional-lethal mutations
temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation
the organisms can the organisms can replicate replicate at a at a
relativelyrelatively low low permissive temperature permissive temperature
but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive
temperaturetemperature
Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize
d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost
errdquo injectionerrdquo injection
A Live (attenuated) A Live (attenuated) vaccinesvaccines
B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate
responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe
toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves
artificial passive artificial passive immunization immunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
Section 2 virology diagnosisSection 2 virology diagnosis
ViralViral infeinfectionction
Electron microscopeElectron microscope
Light microscope inLight microscope inclusion bodyclusion body
isolationisolation
VirusVirus particleparticle
V AgV Ag
V nucleic acidV nucleic acid
Viral enzymeViral enzyme retroviridaseretroviridase
DetectDetect virusvirus
DetectDetect AbAb
Choose the appropriate specimenChoose the appropriate specimen
Aseptic operationAseptic operation add antibiotics to speciadd antibiotics to speci
menmen
Take it in acute phase Take it in acute phase
Low temperature conservationLow temperature conservation transport it transport it
promptlypromptly preserve in 50preserve in 50 glycerineglycerine
Serologial diagnose paired serumSerologial diagnose paired serum
Specimen Selection Collection and ProcessingSpecimen Selection Collection and Processing
二二 viral isolation and identifyingviral isolation and identifying
specimenspecimenAnimal inoculaAnimal inoculationtion
Embryonated eEmbryonated egggg
Cell cultureCell culture
positivepositive
negativenegative
identifyingidentifying
BlindBlind passagepassage22 - 3generatiogenerationn
negativenegative
Virus negatiVirus negativeve
asepsisasepsis
(一)(一) viral isolationviral isolation
11 animal inoculationanimal inoculation
chimpanzeemonkeyrabbitratchimpanzeemonkeyrabbitrat
22 embryonated egg inoculationembryonated egg inoculation
3 Cell culture3 Cell culture (( commonly usedcommonly used ))Primary and secondary cell culturePrimary and secondary cell culture
general proceduregeneral procedure
tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell
cell monolayercell monolayer
brokenbroken proteaseprotease
(primary cell)(primary cell)Serial subcultivationSerial subcultivation
Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)
features of virus cell culturefeatures of virus cell culture
A sensitive to many kinds of virusesA sensitive to many kinds of viruses
B cost highB cost high
C carry latent virusC carry latent virus
(2)Diploid(2)Diploid cellcell cultureculture
usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine
(3) secondary cell culture(3) secondary cell culture
sensitive to sensitive to multiplemultiple viruses viruses
high reproductive capacity long genera high reproductive capacity long generation timetion time
have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine
(( 二二 )) indexindex for reproduction of virusfor reproduction of virus
(1)Virus1)Virus infected cellinfected cell
(( cell roundinggathercell roundinggather
necrosisnecrosis ))
1 Cytopathic effect CPE)1 Cytopathic effect CPE)
CPECPE
(( 22 )) cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation
(( 33 )) viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell
CPECPE
22 hemadsorption hemadsorption HAd HAd
hemagglutininhemagglutinin (( HAHA )) on membrane of vion membrane of virus infected cell rus infected cell
Can Can adsorbadsorb vertebrate vertebrate RBCRBC
3 Interference3 Interference
4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange
( 三 ) viral quantitative assayHemagglutination test total amount
Plaque bacteriophage determination
ID50 or TCID50
Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le
sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex
cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a
nd hyphand hypha
22 cultureculture
specimen sabouraud mediumspecimen sabouraud medium
macroculture microculturemacroculture microculture
colonycolony
biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique
sporespore
3animal test3animal test
4skin test hypersensitivy4skin test hypersensitivy
(三)(三) mycology rapid diagnosismycology rapid diagnosis
serological test detect Ag or Abserological test detect Ag or Ab
detect nucleic acid of fungusdetect nucleic acid of fungus
detect mycotoxindetect mycotoxin
questionsquestions
11 definitiondefinition
serological identificationserological identification
and diagnosis)and diagnosis)
Cytopathic effect CPECytopathic effect CPE
hemadsorption HAdhemadsorption HAd
interferenceinterference
22 principle of Widalrsquos testprinciple of Widalrsquos test
33 principle of Gramrsquos stainingprinciple of Gramrsquos staining
Section 4Section 4
artificial active artificial active immunityimmunity
Artificial active immunityArtificial active immunity
stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid
vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA
A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow
th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni
citycity are repeatedly selected are repeatedly selected BCGBCG
The mutation is expressed only The mutation is expressed only
under certain conditionsunder certain conditions
may be may be useful in vaccinesuseful in vaccines
Conditional-lethal mutationsConditional-lethal mutations
temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation
the organisms can the organisms can replicate replicate at a at a
relativelyrelatively low low permissive temperature permissive temperature
but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive
temperaturetemperature
Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize
d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost
errdquo injectionerrdquo injection
A Live (attenuated) A Live (attenuated) vaccinesvaccines
B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate
responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe
toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves
artificial passive artificial passive immunization immunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
Choose the appropriate specimenChoose the appropriate specimen
Aseptic operationAseptic operation add antibiotics to speciadd antibiotics to speci
menmen
Take it in acute phase Take it in acute phase
Low temperature conservationLow temperature conservation transport it transport it
promptlypromptly preserve in 50preserve in 50 glycerineglycerine
Serologial diagnose paired serumSerologial diagnose paired serum
Specimen Selection Collection and ProcessingSpecimen Selection Collection and Processing
二二 viral isolation and identifyingviral isolation and identifying
specimenspecimenAnimal inoculaAnimal inoculationtion
Embryonated eEmbryonated egggg
Cell cultureCell culture
positivepositive
negativenegative
identifyingidentifying
BlindBlind passagepassage22 - 3generatiogenerationn
negativenegative
Virus negatiVirus negativeve
asepsisasepsis
(一)(一) viral isolationviral isolation
11 animal inoculationanimal inoculation
chimpanzeemonkeyrabbitratchimpanzeemonkeyrabbitrat
22 embryonated egg inoculationembryonated egg inoculation
3 Cell culture3 Cell culture (( commonly usedcommonly used ))Primary and secondary cell culturePrimary and secondary cell culture
general proceduregeneral procedure
tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell
cell monolayercell monolayer
brokenbroken proteaseprotease
(primary cell)(primary cell)Serial subcultivationSerial subcultivation
Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)
features of virus cell culturefeatures of virus cell culture
A sensitive to many kinds of virusesA sensitive to many kinds of viruses
B cost highB cost high
C carry latent virusC carry latent virus
(2)Diploid(2)Diploid cellcell cultureculture
usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine
(3) secondary cell culture(3) secondary cell culture
sensitive to sensitive to multiplemultiple viruses viruses
high reproductive capacity long genera high reproductive capacity long generation timetion time
have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine
(( 二二 )) indexindex for reproduction of virusfor reproduction of virus
(1)Virus1)Virus infected cellinfected cell
(( cell roundinggathercell roundinggather
necrosisnecrosis ))
1 Cytopathic effect CPE)1 Cytopathic effect CPE)
CPECPE
(( 22 )) cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation
(( 33 )) viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell
CPECPE
22 hemadsorption hemadsorption HAd HAd
hemagglutininhemagglutinin (( HAHA )) on membrane of vion membrane of virus infected cell rus infected cell
Can Can adsorbadsorb vertebrate vertebrate RBCRBC
3 Interference3 Interference
4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange
( 三 ) viral quantitative assayHemagglutination test total amount
Plaque bacteriophage determination
ID50 or TCID50
Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le
sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex
cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a
nd hyphand hypha
22 cultureculture
specimen sabouraud mediumspecimen sabouraud medium
macroculture microculturemacroculture microculture
colonycolony
biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique
sporespore
3animal test3animal test
4skin test hypersensitivy4skin test hypersensitivy
(三)(三) mycology rapid diagnosismycology rapid diagnosis
serological test detect Ag or Abserological test detect Ag or Ab
detect nucleic acid of fungusdetect nucleic acid of fungus
detect mycotoxindetect mycotoxin
questionsquestions
11 definitiondefinition
serological identificationserological identification
and diagnosis)and diagnosis)
Cytopathic effect CPECytopathic effect CPE
hemadsorption HAdhemadsorption HAd
interferenceinterference
22 principle of Widalrsquos testprinciple of Widalrsquos test
33 principle of Gramrsquos stainingprinciple of Gramrsquos staining
Section 4Section 4
artificial active artificial active immunityimmunity
Artificial active immunityArtificial active immunity
stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid
vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA
A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow
th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni
citycity are repeatedly selected are repeatedly selected BCGBCG
The mutation is expressed only The mutation is expressed only
under certain conditionsunder certain conditions
may be may be useful in vaccinesuseful in vaccines
Conditional-lethal mutationsConditional-lethal mutations
temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation
the organisms can the organisms can replicate replicate at a at a
relativelyrelatively low low permissive temperature permissive temperature
but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive
temperaturetemperature
Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize
d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost
errdquo injectionerrdquo injection
A Live (attenuated) A Live (attenuated) vaccinesvaccines
B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate
responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe
toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves
artificial passive artificial passive immunization immunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
二二 viral isolation and identifyingviral isolation and identifying
specimenspecimenAnimal inoculaAnimal inoculationtion
Embryonated eEmbryonated egggg
Cell cultureCell culture
positivepositive
negativenegative
identifyingidentifying
BlindBlind passagepassage22 - 3generatiogenerationn
negativenegative
Virus negatiVirus negativeve
asepsisasepsis
(一)(一) viral isolationviral isolation
11 animal inoculationanimal inoculation
chimpanzeemonkeyrabbitratchimpanzeemonkeyrabbitrat
22 embryonated egg inoculationembryonated egg inoculation
3 Cell culture3 Cell culture (( commonly usedcommonly used ))Primary and secondary cell culturePrimary and secondary cell culture
general proceduregeneral procedure
tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell
cell monolayercell monolayer
brokenbroken proteaseprotease
(primary cell)(primary cell)Serial subcultivationSerial subcultivation
Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)
features of virus cell culturefeatures of virus cell culture
A sensitive to many kinds of virusesA sensitive to many kinds of viruses
B cost highB cost high
C carry latent virusC carry latent virus
(2)Diploid(2)Diploid cellcell cultureculture
usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine
(3) secondary cell culture(3) secondary cell culture
sensitive to sensitive to multiplemultiple viruses viruses
high reproductive capacity long genera high reproductive capacity long generation timetion time
have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine
(( 二二 )) indexindex for reproduction of virusfor reproduction of virus
(1)Virus1)Virus infected cellinfected cell
(( cell roundinggathercell roundinggather
necrosisnecrosis ))
1 Cytopathic effect CPE)1 Cytopathic effect CPE)
CPECPE
(( 22 )) cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation
(( 33 )) viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell
CPECPE
22 hemadsorption hemadsorption HAd HAd
hemagglutininhemagglutinin (( HAHA )) on membrane of vion membrane of virus infected cell rus infected cell
Can Can adsorbadsorb vertebrate vertebrate RBCRBC
3 Interference3 Interference
4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange
( 三 ) viral quantitative assayHemagglutination test total amount
Plaque bacteriophage determination
ID50 or TCID50
Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le
sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex
cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a
nd hyphand hypha
22 cultureculture
specimen sabouraud mediumspecimen sabouraud medium
macroculture microculturemacroculture microculture
colonycolony
biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique
sporespore
3animal test3animal test
4skin test hypersensitivy4skin test hypersensitivy
(三)(三) mycology rapid diagnosismycology rapid diagnosis
serological test detect Ag or Abserological test detect Ag or Ab
detect nucleic acid of fungusdetect nucleic acid of fungus
detect mycotoxindetect mycotoxin
questionsquestions
11 definitiondefinition
serological identificationserological identification
and diagnosis)and diagnosis)
Cytopathic effect CPECytopathic effect CPE
hemadsorption HAdhemadsorption HAd
interferenceinterference
22 principle of Widalrsquos testprinciple of Widalrsquos test
33 principle of Gramrsquos stainingprinciple of Gramrsquos staining
Section 4Section 4
artificial active artificial active immunityimmunity
Artificial active immunityArtificial active immunity
stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid
vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA
A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow
th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni
citycity are repeatedly selected are repeatedly selected BCGBCG
The mutation is expressed only The mutation is expressed only
under certain conditionsunder certain conditions
may be may be useful in vaccinesuseful in vaccines
Conditional-lethal mutationsConditional-lethal mutations
temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation
the organisms can the organisms can replicate replicate at a at a
relativelyrelatively low low permissive temperature permissive temperature
but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive
temperaturetemperature
Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize
d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost
errdquo injectionerrdquo injection
A Live (attenuated) A Live (attenuated) vaccinesvaccines
B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate
responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe
toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves
artificial passive artificial passive immunization immunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
(一)(一) viral isolationviral isolation
11 animal inoculationanimal inoculation
chimpanzeemonkeyrabbitratchimpanzeemonkeyrabbitrat
22 embryonated egg inoculationembryonated egg inoculation
3 Cell culture3 Cell culture (( commonly usedcommonly used ))Primary and secondary cell culturePrimary and secondary cell culture
general proceduregeneral procedure
tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell
cell monolayercell monolayer
brokenbroken proteaseprotease
(primary cell)(primary cell)Serial subcultivationSerial subcultivation
Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)
features of virus cell culturefeatures of virus cell culture
A sensitive to many kinds of virusesA sensitive to many kinds of viruses
B cost highB cost high
C carry latent virusC carry latent virus
(2)Diploid(2)Diploid cellcell cultureculture
usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine
(3) secondary cell culture(3) secondary cell culture
sensitive to sensitive to multiplemultiple viruses viruses
high reproductive capacity long genera high reproductive capacity long generation timetion time
have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine
(( 二二 )) indexindex for reproduction of virusfor reproduction of virus
(1)Virus1)Virus infected cellinfected cell
(( cell roundinggathercell roundinggather
necrosisnecrosis ))
1 Cytopathic effect CPE)1 Cytopathic effect CPE)
CPECPE
(( 22 )) cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation
(( 33 )) viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell
CPECPE
22 hemadsorption hemadsorption HAd HAd
hemagglutininhemagglutinin (( HAHA )) on membrane of vion membrane of virus infected cell rus infected cell
Can Can adsorbadsorb vertebrate vertebrate RBCRBC
3 Interference3 Interference
4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange
( 三 ) viral quantitative assayHemagglutination test total amount
Plaque bacteriophage determination
ID50 or TCID50
Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le
sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex
cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a
nd hyphand hypha
22 cultureculture
specimen sabouraud mediumspecimen sabouraud medium
macroculture microculturemacroculture microculture
colonycolony
biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique
sporespore
3animal test3animal test
4skin test hypersensitivy4skin test hypersensitivy
(三)(三) mycology rapid diagnosismycology rapid diagnosis
serological test detect Ag or Abserological test detect Ag or Ab
detect nucleic acid of fungusdetect nucleic acid of fungus
detect mycotoxindetect mycotoxin
questionsquestions
11 definitiondefinition
serological identificationserological identification
and diagnosis)and diagnosis)
Cytopathic effect CPECytopathic effect CPE
hemadsorption HAdhemadsorption HAd
interferenceinterference
22 principle of Widalrsquos testprinciple of Widalrsquos test
33 principle of Gramrsquos stainingprinciple of Gramrsquos staining
Section 4Section 4
artificial active artificial active immunityimmunity
Artificial active immunityArtificial active immunity
stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid
vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA
A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow
th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni
citycity are repeatedly selected are repeatedly selected BCGBCG
The mutation is expressed only The mutation is expressed only
under certain conditionsunder certain conditions
may be may be useful in vaccinesuseful in vaccines
Conditional-lethal mutationsConditional-lethal mutations
temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation
the organisms can the organisms can replicate replicate at a at a
relativelyrelatively low low permissive temperature permissive temperature
but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive
temperaturetemperature
Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize
d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost
errdquo injectionerrdquo injection
A Live (attenuated) A Live (attenuated) vaccinesvaccines
B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate
responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe
toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves
artificial passive artificial passive immunization immunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
22 embryonated egg inoculationembryonated egg inoculation
3 Cell culture3 Cell culture (( commonly usedcommonly used ))Primary and secondary cell culturePrimary and secondary cell culture
general proceduregeneral procedure
tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell
cell monolayercell monolayer
brokenbroken proteaseprotease
(primary cell)(primary cell)Serial subcultivationSerial subcultivation
Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)
features of virus cell culturefeatures of virus cell culture
A sensitive to many kinds of virusesA sensitive to many kinds of viruses
B cost highB cost high
C carry latent virusC carry latent virus
(2)Diploid(2)Diploid cellcell cultureculture
usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine
(3) secondary cell culture(3) secondary cell culture
sensitive to sensitive to multiplemultiple viruses viruses
high reproductive capacity long genera high reproductive capacity long generation timetion time
have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine
(( 二二 )) indexindex for reproduction of virusfor reproduction of virus
(1)Virus1)Virus infected cellinfected cell
(( cell roundinggathercell roundinggather
necrosisnecrosis ))
1 Cytopathic effect CPE)1 Cytopathic effect CPE)
CPECPE
(( 22 )) cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation
(( 33 )) viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell
CPECPE
22 hemadsorption hemadsorption HAd HAd
hemagglutininhemagglutinin (( HAHA )) on membrane of vion membrane of virus infected cell rus infected cell
Can Can adsorbadsorb vertebrate vertebrate RBCRBC
3 Interference3 Interference
4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange
( 三 ) viral quantitative assayHemagglutination test total amount
Plaque bacteriophage determination
ID50 or TCID50
Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le
sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex
cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a
nd hyphand hypha
22 cultureculture
specimen sabouraud mediumspecimen sabouraud medium
macroculture microculturemacroculture microculture
colonycolony
biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique
sporespore
3animal test3animal test
4skin test hypersensitivy4skin test hypersensitivy
(三)(三) mycology rapid diagnosismycology rapid diagnosis
serological test detect Ag or Abserological test detect Ag or Ab
detect nucleic acid of fungusdetect nucleic acid of fungus
detect mycotoxindetect mycotoxin
questionsquestions
11 definitiondefinition
serological identificationserological identification
and diagnosis)and diagnosis)
Cytopathic effect CPECytopathic effect CPE
hemadsorption HAdhemadsorption HAd
interferenceinterference
22 principle of Widalrsquos testprinciple of Widalrsquos test
33 principle of Gramrsquos stainingprinciple of Gramrsquos staining
Section 4Section 4
artificial active artificial active immunityimmunity
Artificial active immunityArtificial active immunity
stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid
vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA
A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow
th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni
citycity are repeatedly selected are repeatedly selected BCGBCG
The mutation is expressed only The mutation is expressed only
under certain conditionsunder certain conditions
may be may be useful in vaccinesuseful in vaccines
Conditional-lethal mutationsConditional-lethal mutations
temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation
the organisms can the organisms can replicate replicate at a at a
relativelyrelatively low low permissive temperature permissive temperature
but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive
temperaturetemperature
Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize
d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost
errdquo injectionerrdquo injection
A Live (attenuated) A Live (attenuated) vaccinesvaccines
B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate
responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe
toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves
artificial passive artificial passive immunization immunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
3 Cell culture3 Cell culture (( commonly usedcommonly used ))Primary and secondary cell culturePrimary and secondary cell culture
general proceduregeneral procedure
tissuetissue tissuetissue piecepiece sporadicsporadic singlesingle cellcell
cell monolayercell monolayer
brokenbroken proteaseprotease
(primary cell)(primary cell)Serial subcultivationSerial subcultivation
Cell monolayerCell monolayer(secondary cell (secondary cell culture)culture)
features of virus cell culturefeatures of virus cell culture
A sensitive to many kinds of virusesA sensitive to many kinds of viruses
B cost highB cost high
C carry latent virusC carry latent virus
(2)Diploid(2)Diploid cellcell cultureculture
usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine
(3) secondary cell culture(3) secondary cell culture
sensitive to sensitive to multiplemultiple viruses viruses
high reproductive capacity long genera high reproductive capacity long generation timetion time
have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine
(( 二二 )) indexindex for reproduction of virusfor reproduction of virus
(1)Virus1)Virus infected cellinfected cell
(( cell roundinggathercell roundinggather
necrosisnecrosis ))
1 Cytopathic effect CPE)1 Cytopathic effect CPE)
CPECPE
(( 22 )) cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation
(( 33 )) viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell
CPECPE
22 hemadsorption hemadsorption HAd HAd
hemagglutininhemagglutinin (( HAHA )) on membrane of vion membrane of virus infected cell rus infected cell
Can Can adsorbadsorb vertebrate vertebrate RBCRBC
3 Interference3 Interference
4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange
( 三 ) viral quantitative assayHemagglutination test total amount
Plaque bacteriophage determination
ID50 or TCID50
Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le
sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex
cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a
nd hyphand hypha
22 cultureculture
specimen sabouraud mediumspecimen sabouraud medium
macroculture microculturemacroculture microculture
colonycolony
biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique
sporespore
3animal test3animal test
4skin test hypersensitivy4skin test hypersensitivy
(三)(三) mycology rapid diagnosismycology rapid diagnosis
serological test detect Ag or Abserological test detect Ag or Ab
detect nucleic acid of fungusdetect nucleic acid of fungus
detect mycotoxindetect mycotoxin
questionsquestions
11 definitiondefinition
serological identificationserological identification
and diagnosis)and diagnosis)
Cytopathic effect CPECytopathic effect CPE
hemadsorption HAdhemadsorption HAd
interferenceinterference
22 principle of Widalrsquos testprinciple of Widalrsquos test
33 principle of Gramrsquos stainingprinciple of Gramrsquos staining
Section 4Section 4
artificial active artificial active immunityimmunity
Artificial active immunityArtificial active immunity
stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid
vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA
A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow
th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni
citycity are repeatedly selected are repeatedly selected BCGBCG
The mutation is expressed only The mutation is expressed only
under certain conditionsunder certain conditions
may be may be useful in vaccinesuseful in vaccines
Conditional-lethal mutationsConditional-lethal mutations
temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation
the organisms can the organisms can replicate replicate at a at a
relativelyrelatively low low permissive temperature permissive temperature
but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive
temperaturetemperature
Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize
d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost
errdquo injectionerrdquo injection
A Live (attenuated) A Live (attenuated) vaccinesvaccines
B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate
responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe
toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves
artificial passive artificial passive immunization immunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
features of virus cell culturefeatures of virus cell culture
A sensitive to many kinds of virusesA sensitive to many kinds of viruses
B cost highB cost high
C carry latent virusC carry latent virus
(2)Diploid(2)Diploid cellcell cultureculture
usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine
(3) secondary cell culture(3) secondary cell culture
sensitive to sensitive to multiplemultiple viruses viruses
high reproductive capacity long genera high reproductive capacity long generation timetion time
have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine
(( 二二 )) indexindex for reproduction of virusfor reproduction of virus
(1)Virus1)Virus infected cellinfected cell
(( cell roundinggathercell roundinggather
necrosisnecrosis ))
1 Cytopathic effect CPE)1 Cytopathic effect CPE)
CPECPE
(( 22 )) cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation
(( 33 )) viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell
CPECPE
22 hemadsorption hemadsorption HAd HAd
hemagglutininhemagglutinin (( HAHA )) on membrane of vion membrane of virus infected cell rus infected cell
Can Can adsorbadsorb vertebrate vertebrate RBCRBC
3 Interference3 Interference
4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange
( 三 ) viral quantitative assayHemagglutination test total amount
Plaque bacteriophage determination
ID50 or TCID50
Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le
sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex
cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a
nd hyphand hypha
22 cultureculture
specimen sabouraud mediumspecimen sabouraud medium
macroculture microculturemacroculture microculture
colonycolony
biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique
sporespore
3animal test3animal test
4skin test hypersensitivy4skin test hypersensitivy
(三)(三) mycology rapid diagnosismycology rapid diagnosis
serological test detect Ag or Abserological test detect Ag or Ab
detect nucleic acid of fungusdetect nucleic acid of fungus
detect mycotoxindetect mycotoxin
questionsquestions
11 definitiondefinition
serological identificationserological identification
and diagnosis)and diagnosis)
Cytopathic effect CPECytopathic effect CPE
hemadsorption HAdhemadsorption HAd
interferenceinterference
22 principle of Widalrsquos testprinciple of Widalrsquos test
33 principle of Gramrsquos stainingprinciple of Gramrsquos staining
Section 4Section 4
artificial active artificial active immunityimmunity
Artificial active immunityArtificial active immunity
stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid
vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA
A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow
th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni
citycity are repeatedly selected are repeatedly selected BCGBCG
The mutation is expressed only The mutation is expressed only
under certain conditionsunder certain conditions
may be may be useful in vaccinesuseful in vaccines
Conditional-lethal mutationsConditional-lethal mutations
temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation
the organisms can the organisms can replicate replicate at a at a
relativelyrelatively low low permissive temperature permissive temperature
but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive
temperaturetemperature
Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize
d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost
errdquo injectionerrdquo injection
A Live (attenuated) A Live (attenuated) vaccinesvaccines
B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate
responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe
toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves
artificial passive artificial passive immunization immunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
(2)Diploid(2)Diploid cellcell cultureculture
usedused to isolate virus and to isolate virus and obtain obtain vaccinevaccine
(3) secondary cell culture(3) secondary cell culture
sensitive to sensitive to multiplemultiple viruses viruses
high reproductive capacity long genera high reproductive capacity long generation timetion time
have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine
(( 二二 )) indexindex for reproduction of virusfor reproduction of virus
(1)Virus1)Virus infected cellinfected cell
(( cell roundinggathercell roundinggather
necrosisnecrosis ))
1 Cytopathic effect CPE)1 Cytopathic effect CPE)
CPECPE
(( 22 )) cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation
(( 33 )) viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell
CPECPE
22 hemadsorption hemadsorption HAd HAd
hemagglutininhemagglutinin (( HAHA )) on membrane of vion membrane of virus infected cell rus infected cell
Can Can adsorbadsorb vertebrate vertebrate RBCRBC
3 Interference3 Interference
4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange
( 三 ) viral quantitative assayHemagglutination test total amount
Plaque bacteriophage determination
ID50 or TCID50
Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le
sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex
cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a
nd hyphand hypha
22 cultureculture
specimen sabouraud mediumspecimen sabouraud medium
macroculture microculturemacroculture microculture
colonycolony
biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique
sporespore
3animal test3animal test
4skin test hypersensitivy4skin test hypersensitivy
(三)(三) mycology rapid diagnosismycology rapid diagnosis
serological test detect Ag or Abserological test detect Ag or Ab
detect nucleic acid of fungusdetect nucleic acid of fungus
detect mycotoxindetect mycotoxin
questionsquestions
11 definitiondefinition
serological identificationserological identification
and diagnosis)and diagnosis)
Cytopathic effect CPECytopathic effect CPE
hemadsorption HAdhemadsorption HAd
interferenceinterference
22 principle of Widalrsquos testprinciple of Widalrsquos test
33 principle of Gramrsquos stainingprinciple of Gramrsquos staining
Section 4Section 4
artificial active artificial active immunityimmunity
Artificial active immunityArtificial active immunity
stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid
vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA
A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow
th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni
citycity are repeatedly selected are repeatedly selected BCGBCG
The mutation is expressed only The mutation is expressed only
under certain conditionsunder certain conditions
may be may be useful in vaccinesuseful in vaccines
Conditional-lethal mutationsConditional-lethal mutations
temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation
the organisms can the organisms can replicate replicate at a at a
relativelyrelatively low low permissive temperature permissive temperature
but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive
temperaturetemperature
Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize
d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost
errdquo injectionerrdquo injection
A Live (attenuated) A Live (attenuated) vaccinesvaccines
B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate
responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe
toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves
artificial passive artificial passive immunization immunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
(3) secondary cell culture(3) secondary cell culture
sensitive to sensitive to multiplemultiple viruses viruses
high reproductive capacity long genera high reproductive capacity long generation timetion time
have have danger of carcinogenesisdanger of carcinogenesis cannocannot use to produce vaccinet use to produce vaccine
(( 二二 )) indexindex for reproduction of virusfor reproduction of virus
(1)Virus1)Virus infected cellinfected cell
(( cell roundinggathercell roundinggather
necrosisnecrosis ))
1 Cytopathic effect CPE)1 Cytopathic effect CPE)
CPECPE
(( 22 )) cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation
(( 33 )) viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell
CPECPE
22 hemadsorption hemadsorption HAd HAd
hemagglutininhemagglutinin (( HAHA )) on membrane of vion membrane of virus infected cell rus infected cell
Can Can adsorbadsorb vertebrate vertebrate RBCRBC
3 Interference3 Interference
4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange
( 三 ) viral quantitative assayHemagglutination test total amount
Plaque bacteriophage determination
ID50 or TCID50
Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le
sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex
cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a
nd hyphand hypha
22 cultureculture
specimen sabouraud mediumspecimen sabouraud medium
macroculture microculturemacroculture microculture
colonycolony
biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique
sporespore
3animal test3animal test
4skin test hypersensitivy4skin test hypersensitivy
(三)(三) mycology rapid diagnosismycology rapid diagnosis
serological test detect Ag or Abserological test detect Ag or Ab
detect nucleic acid of fungusdetect nucleic acid of fungus
detect mycotoxindetect mycotoxin
questionsquestions
11 definitiondefinition
serological identificationserological identification
and diagnosis)and diagnosis)
Cytopathic effect CPECytopathic effect CPE
hemadsorption HAdhemadsorption HAd
interferenceinterference
22 principle of Widalrsquos testprinciple of Widalrsquos test
33 principle of Gramrsquos stainingprinciple of Gramrsquos staining
Section 4Section 4
artificial active artificial active immunityimmunity
Artificial active immunityArtificial active immunity
stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid
vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA
A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow
th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni
citycity are repeatedly selected are repeatedly selected BCGBCG
The mutation is expressed only The mutation is expressed only
under certain conditionsunder certain conditions
may be may be useful in vaccinesuseful in vaccines
Conditional-lethal mutationsConditional-lethal mutations
temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation
the organisms can the organisms can replicate replicate at a at a
relativelyrelatively low low permissive temperature permissive temperature
but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive
temperaturetemperature
Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize
d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost
errdquo injectionerrdquo injection
A Live (attenuated) A Live (attenuated) vaccinesvaccines
B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate
responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe
toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves
artificial passive artificial passive immunization immunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
(( 二二 )) indexindex for reproduction of virusfor reproduction of virus
(1)Virus1)Virus infected cellinfected cell
(( cell roundinggathercell roundinggather
necrosisnecrosis ))
1 Cytopathic effect CPE)1 Cytopathic effect CPE)
CPECPE
(( 22 )) cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation
(( 33 )) viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell
CPECPE
22 hemadsorption hemadsorption HAd HAd
hemagglutininhemagglutinin (( HAHA )) on membrane of vion membrane of virus infected cell rus infected cell
Can Can adsorbadsorb vertebrate vertebrate RBCRBC
3 Interference3 Interference
4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange
( 三 ) viral quantitative assayHemagglutination test total amount
Plaque bacteriophage determination
ID50 or TCID50
Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le
sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex
cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a
nd hyphand hypha
22 cultureculture
specimen sabouraud mediumspecimen sabouraud medium
macroculture microculturemacroculture microculture
colonycolony
biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique
sporespore
3animal test3animal test
4skin test hypersensitivy4skin test hypersensitivy
(三)(三) mycology rapid diagnosismycology rapid diagnosis
serological test detect Ag or Abserological test detect Ag or Ab
detect nucleic acid of fungusdetect nucleic acid of fungus
detect mycotoxindetect mycotoxin
questionsquestions
11 definitiondefinition
serological identificationserological identification
and diagnosis)and diagnosis)
Cytopathic effect CPECytopathic effect CPE
hemadsorption HAdhemadsorption HAd
interferenceinterference
22 principle of Widalrsquos testprinciple of Widalrsquos test
33 principle of Gramrsquos stainingprinciple of Gramrsquos staining
Section 4Section 4
artificial active artificial active immunityimmunity
Artificial active immunityArtificial active immunity
stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid
vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA
A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow
th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni
citycity are repeatedly selected are repeatedly selected BCGBCG
The mutation is expressed only The mutation is expressed only
under certain conditionsunder certain conditions
may be may be useful in vaccinesuseful in vaccines
Conditional-lethal mutationsConditional-lethal mutations
temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation
the organisms can the organisms can replicate replicate at a at a
relativelyrelatively low low permissive temperature permissive temperature
but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive
temperaturetemperature
Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize
d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost
errdquo injectionerrdquo injection
A Live (attenuated) A Live (attenuated) vaccinesvaccines
B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate
responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe
toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves
artificial passive artificial passive immunization immunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
CPECPE
(( 22 )) cell fusioncell fusion aft after virus infectioner virus infectionMultinucleated giant Multinucleated giant cell formationcell formation
(( 33 )) viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell
CPECPE
22 hemadsorption hemadsorption HAd HAd
hemagglutininhemagglutinin (( HAHA )) on membrane of vion membrane of virus infected cell rus infected cell
Can Can adsorbadsorb vertebrate vertebrate RBCRBC
3 Interference3 Interference
4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange
( 三 ) viral quantitative assayHemagglutination test total amount
Plaque bacteriophage determination
ID50 or TCID50
Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le
sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex
cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a
nd hyphand hypha
22 cultureculture
specimen sabouraud mediumspecimen sabouraud medium
macroculture microculturemacroculture microculture
colonycolony
biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique
sporespore
3animal test3animal test
4skin test hypersensitivy4skin test hypersensitivy
(三)(三) mycology rapid diagnosismycology rapid diagnosis
serological test detect Ag or Abserological test detect Ag or Ab
detect nucleic acid of fungusdetect nucleic acid of fungus
detect mycotoxindetect mycotoxin
questionsquestions
11 definitiondefinition
serological identificationserological identification
and diagnosis)and diagnosis)
Cytopathic effect CPECytopathic effect CPE
hemadsorption HAdhemadsorption HAd
interferenceinterference
22 principle of Widalrsquos testprinciple of Widalrsquos test
33 principle of Gramrsquos stainingprinciple of Gramrsquos staining
Section 4Section 4
artificial active artificial active immunityimmunity
Artificial active immunityArtificial active immunity
stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid
vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA
A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow
th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni
citycity are repeatedly selected are repeatedly selected BCGBCG
The mutation is expressed only The mutation is expressed only
under certain conditionsunder certain conditions
may be may be useful in vaccinesuseful in vaccines
Conditional-lethal mutationsConditional-lethal mutations
temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation
the organisms can the organisms can replicate replicate at a at a
relativelyrelatively low low permissive temperature permissive temperature
but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive
temperaturetemperature
Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize
d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost
errdquo injectionerrdquo injection
A Live (attenuated) A Live (attenuated) vaccinesvaccines
B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate
responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe
toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves
artificial passive artificial passive immunization immunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
(( 33 )) viral inclusionviral inclusion in cytoplasm or nucin cytoplasm or nucleus of infected cellleus of infected cell
CPECPE
22 hemadsorption hemadsorption HAd HAd
hemagglutininhemagglutinin (( HAHA )) on membrane of vion membrane of virus infected cell rus infected cell
Can Can adsorbadsorb vertebrate vertebrate RBCRBC
3 Interference3 Interference
4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange
( 三 ) viral quantitative assayHemagglutination test total amount
Plaque bacteriophage determination
ID50 or TCID50
Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le
sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex
cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a
nd hyphand hypha
22 cultureculture
specimen sabouraud mediumspecimen sabouraud medium
macroculture microculturemacroculture microculture
colonycolony
biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique
sporespore
3animal test3animal test
4skin test hypersensitivy4skin test hypersensitivy
(三)(三) mycology rapid diagnosismycology rapid diagnosis
serological test detect Ag or Abserological test detect Ag or Ab
detect nucleic acid of fungusdetect nucleic acid of fungus
detect mycotoxindetect mycotoxin
questionsquestions
11 definitiondefinition
serological identificationserological identification
and diagnosis)and diagnosis)
Cytopathic effect CPECytopathic effect CPE
hemadsorption HAdhemadsorption HAd
interferenceinterference
22 principle of Widalrsquos testprinciple of Widalrsquos test
33 principle of Gramrsquos stainingprinciple of Gramrsquos staining
Section 4Section 4
artificial active artificial active immunityimmunity
Artificial active immunityArtificial active immunity
stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid
vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA
A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow
th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni
citycity are repeatedly selected are repeatedly selected BCGBCG
The mutation is expressed only The mutation is expressed only
under certain conditionsunder certain conditions
may be may be useful in vaccinesuseful in vaccines
Conditional-lethal mutationsConditional-lethal mutations
temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation
the organisms can the organisms can replicate replicate at a at a
relativelyrelatively low low permissive temperature permissive temperature
but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive
temperaturetemperature
Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize
d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost
errdquo injectionerrdquo injection
A Live (attenuated) A Live (attenuated) vaccinesvaccines
B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate
responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe
toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves
artificial passive artificial passive immunization immunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
22 hemadsorption hemadsorption HAd HAd
hemagglutininhemagglutinin (( HAHA )) on membrane of vion membrane of virus infected cell rus infected cell
Can Can adsorbadsorb vertebrate vertebrate RBCRBC
3 Interference3 Interference
4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange
( 三 ) viral quantitative assayHemagglutination test total amount
Plaque bacteriophage determination
ID50 or TCID50
Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le
sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex
cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a
nd hyphand hypha
22 cultureculture
specimen sabouraud mediumspecimen sabouraud medium
macroculture microculturemacroculture microculture
colonycolony
biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique
sporespore
3animal test3animal test
4skin test hypersensitivy4skin test hypersensitivy
(三)(三) mycology rapid diagnosismycology rapid diagnosis
serological test detect Ag or Abserological test detect Ag or Ab
detect nucleic acid of fungusdetect nucleic acid of fungus
detect mycotoxindetect mycotoxin
questionsquestions
11 definitiondefinition
serological identificationserological identification
and diagnosis)and diagnosis)
Cytopathic effect CPECytopathic effect CPE
hemadsorption HAdhemadsorption HAd
interferenceinterference
22 principle of Widalrsquos testprinciple of Widalrsquos test
33 principle of Gramrsquos stainingprinciple of Gramrsquos staining
Section 4Section 4
artificial active artificial active immunityimmunity
Artificial active immunityArtificial active immunity
stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid
vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA
A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow
th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni
citycity are repeatedly selected are repeatedly selected BCGBCG
The mutation is expressed only The mutation is expressed only
under certain conditionsunder certain conditions
may be may be useful in vaccinesuseful in vaccines
Conditional-lethal mutationsConditional-lethal mutations
temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation
the organisms can the organisms can replicate replicate at a at a
relativelyrelatively low low permissive temperature permissive temperature
but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive
temperaturetemperature
Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize
d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost
errdquo injectionerrdquo injection
A Live (attenuated) A Live (attenuated) vaccinesvaccines
B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate
responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe
toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves
artificial passive artificial passive immunization immunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
3 Interference3 Interference
4 Cell metabolism change4 Cell metabolism change Virus reproductionVirus reproduction pH pH of culture of culture cchangehange
( 三 ) viral quantitative assayHemagglutination test total amount
Plaque bacteriophage determination
ID50 or TCID50
Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le
sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex
cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a
nd hyphand hypha
22 cultureculture
specimen sabouraud mediumspecimen sabouraud medium
macroculture microculturemacroculture microculture
colonycolony
biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique
sporespore
3animal test3animal test
4skin test hypersensitivy4skin test hypersensitivy
(三)(三) mycology rapid diagnosismycology rapid diagnosis
serological test detect Ag or Abserological test detect Ag or Ab
detect nucleic acid of fungusdetect nucleic acid of fungus
detect mycotoxindetect mycotoxin
questionsquestions
11 definitiondefinition
serological identificationserological identification
and diagnosis)and diagnosis)
Cytopathic effect CPECytopathic effect CPE
hemadsorption HAdhemadsorption HAd
interferenceinterference
22 principle of Widalrsquos testprinciple of Widalrsquos test
33 principle of Gramrsquos stainingprinciple of Gramrsquos staining
Section 4Section 4
artificial active artificial active immunityimmunity
Artificial active immunityArtificial active immunity
stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid
vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA
A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow
th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni
citycity are repeatedly selected are repeatedly selected BCGBCG
The mutation is expressed only The mutation is expressed only
under certain conditionsunder certain conditions
may be may be useful in vaccinesuseful in vaccines
Conditional-lethal mutationsConditional-lethal mutations
temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation
the organisms can the organisms can replicate replicate at a at a
relativelyrelatively low low permissive temperature permissive temperature
but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive
temperaturetemperature
Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize
d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost
errdquo injectionerrdquo injection
A Live (attenuated) A Live (attenuated) vaccinesvaccines
B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate
responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe
toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves
artificial passive artificial passive immunization immunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
( 三 ) viral quantitative assayHemagglutination test total amount
Plaque bacteriophage determination
ID50 or TCID50
Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le
sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex
cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a
nd hyphand hypha
22 cultureculture
specimen sabouraud mediumspecimen sabouraud medium
macroculture microculturemacroculture microculture
colonycolony
biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique
sporespore
3animal test3animal test
4skin test hypersensitivy4skin test hypersensitivy
(三)(三) mycology rapid diagnosismycology rapid diagnosis
serological test detect Ag or Abserological test detect Ag or Ab
detect nucleic acid of fungusdetect nucleic acid of fungus
detect mycotoxindetect mycotoxin
questionsquestions
11 definitiondefinition
serological identificationserological identification
and diagnosis)and diagnosis)
Cytopathic effect CPECytopathic effect CPE
hemadsorption HAdhemadsorption HAd
interferenceinterference
22 principle of Widalrsquos testprinciple of Widalrsquos test
33 principle of Gramrsquos stainingprinciple of Gramrsquos staining
Section 4Section 4
artificial active artificial active immunityimmunity
Artificial active immunityArtificial active immunity
stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid
vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA
A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow
th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni
citycity are repeatedly selected are repeatedly selected BCGBCG
The mutation is expressed only The mutation is expressed only
under certain conditionsunder certain conditions
may be may be useful in vaccinesuseful in vaccines
Conditional-lethal mutationsConditional-lethal mutations
temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation
the organisms can the organisms can replicate replicate at a at a
relativelyrelatively low low permissive temperature permissive temperature
but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive
temperaturetemperature
Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize
d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost
errdquo injectionerrdquo injection
A Live (attenuated) A Live (attenuated) vaccinesvaccines
B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate
responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe
toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves
artificial passive artificial passive immunization immunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
Section 3 mycology diagnosisSection 3 mycology diagnosis(( 一一 ) collection of specimens) collection of specimensSuperficial infected fungusSuperficial infected fungus surface layer skin lesurface layer skin le
sion tissuesion tissue deep infected fungusdeep infected fungus body fluid secretion exbody fluid secretion ex
cretioncretion(( 二二 )detection and identify)detection and identify11 direct observation with microscopedirect observation with microscope spore aspore a
nd hyphand hypha
22 cultureculture
specimen sabouraud mediumspecimen sabouraud medium
macroculture microculturemacroculture microculture
colonycolony
biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique
sporespore
3animal test3animal test
4skin test hypersensitivy4skin test hypersensitivy
(三)(三) mycology rapid diagnosismycology rapid diagnosis
serological test detect Ag or Abserological test detect Ag or Ab
detect nucleic acid of fungusdetect nucleic acid of fungus
detect mycotoxindetect mycotoxin
questionsquestions
11 definitiondefinition
serological identificationserological identification
and diagnosis)and diagnosis)
Cytopathic effect CPECytopathic effect CPE
hemadsorption HAdhemadsorption HAd
interferenceinterference
22 principle of Widalrsquos testprinciple of Widalrsquos test
33 principle of Gramrsquos stainingprinciple of Gramrsquos staining
Section 4Section 4
artificial active artificial active immunityimmunity
Artificial active immunityArtificial active immunity
stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid
vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA
A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow
th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni
citycity are repeatedly selected are repeatedly selected BCGBCG
The mutation is expressed only The mutation is expressed only
under certain conditionsunder certain conditions
may be may be useful in vaccinesuseful in vaccines
Conditional-lethal mutationsConditional-lethal mutations
temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation
the organisms can the organisms can replicate replicate at a at a
relativelyrelatively low low permissive temperature permissive temperature
but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive
temperaturetemperature
Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize
d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost
errdquo injectionerrdquo injection
A Live (attenuated) A Live (attenuated) vaccinesvaccines
B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate
responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe
toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves
artificial passive artificial passive immunization immunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
22 cultureculture
specimen sabouraud mediumspecimen sabouraud medium
macroculture microculturemacroculture microculture
colonycolony
biochemical reaction observe biochemical reaction observe or hyphaor hypha molecular biologymolecular biology techniquetechnique
sporespore
3animal test3animal test
4skin test hypersensitivy4skin test hypersensitivy
(三)(三) mycology rapid diagnosismycology rapid diagnosis
serological test detect Ag or Abserological test detect Ag or Ab
detect nucleic acid of fungusdetect nucleic acid of fungus
detect mycotoxindetect mycotoxin
questionsquestions
11 definitiondefinition
serological identificationserological identification
and diagnosis)and diagnosis)
Cytopathic effect CPECytopathic effect CPE
hemadsorption HAdhemadsorption HAd
interferenceinterference
22 principle of Widalrsquos testprinciple of Widalrsquos test
33 principle of Gramrsquos stainingprinciple of Gramrsquos staining
Section 4Section 4
artificial active artificial active immunityimmunity
Artificial active immunityArtificial active immunity
stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid
vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA
A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow
th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni
citycity are repeatedly selected are repeatedly selected BCGBCG
The mutation is expressed only The mutation is expressed only
under certain conditionsunder certain conditions
may be may be useful in vaccinesuseful in vaccines
Conditional-lethal mutationsConditional-lethal mutations
temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation
the organisms can the organisms can replicate replicate at a at a
relativelyrelatively low low permissive temperature permissive temperature
but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive
temperaturetemperature
Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize
d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost
errdquo injectionerrdquo injection
A Live (attenuated) A Live (attenuated) vaccinesvaccines
B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate
responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe
toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves
artificial passive artificial passive immunization immunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
3animal test3animal test
4skin test hypersensitivy4skin test hypersensitivy
(三)(三) mycology rapid diagnosismycology rapid diagnosis
serological test detect Ag or Abserological test detect Ag or Ab
detect nucleic acid of fungusdetect nucleic acid of fungus
detect mycotoxindetect mycotoxin
questionsquestions
11 definitiondefinition
serological identificationserological identification
and diagnosis)and diagnosis)
Cytopathic effect CPECytopathic effect CPE
hemadsorption HAdhemadsorption HAd
interferenceinterference
22 principle of Widalrsquos testprinciple of Widalrsquos test
33 principle of Gramrsquos stainingprinciple of Gramrsquos staining
Section 4Section 4
artificial active artificial active immunityimmunity
Artificial active immunityArtificial active immunity
stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid
vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA
A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow
th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni
citycity are repeatedly selected are repeatedly selected BCGBCG
The mutation is expressed only The mutation is expressed only
under certain conditionsunder certain conditions
may be may be useful in vaccinesuseful in vaccines
Conditional-lethal mutationsConditional-lethal mutations
temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation
the organisms can the organisms can replicate replicate at a at a
relativelyrelatively low low permissive temperature permissive temperature
but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive
temperaturetemperature
Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize
d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost
errdquo injectionerrdquo injection
A Live (attenuated) A Live (attenuated) vaccinesvaccines
B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate
responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe
toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves
artificial passive artificial passive immunization immunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
questionsquestions
11 definitiondefinition
serological identificationserological identification
and diagnosis)and diagnosis)
Cytopathic effect CPECytopathic effect CPE
hemadsorption HAdhemadsorption HAd
interferenceinterference
22 principle of Widalrsquos testprinciple of Widalrsquos test
33 principle of Gramrsquos stainingprinciple of Gramrsquos staining
Section 4Section 4
artificial active artificial active immunityimmunity
Artificial active immunityArtificial active immunity
stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid
vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA
A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow
th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni
citycity are repeatedly selected are repeatedly selected BCGBCG
The mutation is expressed only The mutation is expressed only
under certain conditionsunder certain conditions
may be may be useful in vaccinesuseful in vaccines
Conditional-lethal mutationsConditional-lethal mutations
temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation
the organisms can the organisms can replicate replicate at a at a
relativelyrelatively low low permissive temperature permissive temperature
but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive
temperaturetemperature
Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize
d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost
errdquo injectionerrdquo injection
A Live (attenuated) A Live (attenuated) vaccinesvaccines
B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate
responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe
toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves
artificial passive artificial passive immunization immunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
Section 4Section 4
artificial active artificial active immunityimmunity
Artificial active immunityArtificial active immunity
stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid
vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA
A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow
th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni
citycity are repeatedly selected are repeatedly selected BCGBCG
The mutation is expressed only The mutation is expressed only
under certain conditionsunder certain conditions
may be may be useful in vaccinesuseful in vaccines
Conditional-lethal mutationsConditional-lethal mutations
temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation
the organisms can the organisms can replicate replicate at a at a
relativelyrelatively low low permissive temperature permissive temperature
but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive
temperaturetemperature
Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize
d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost
errdquo injectionerrdquo injection
A Live (attenuated) A Live (attenuated) vaccinesvaccines
B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate
responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe
toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves
artificial passive artificial passive immunization immunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
Artificial active immunityArtificial active immunity
stimulate the bodyrsquos immune mecstimulate the bodyrsquos immune mechanisms through administration of a hanisms through administration of a vaccine or toxoidvaccine or toxoid
vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA
A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow
th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni
citycity are repeatedly selected are repeatedly selected BCGBCG
The mutation is expressed only The mutation is expressed only
under certain conditionsunder certain conditions
may be may be useful in vaccinesuseful in vaccines
Conditional-lethal mutationsConditional-lethal mutations
temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation
the organisms can the organisms can replicate replicate at a at a
relativelyrelatively low low permissive temperature permissive temperature
but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive
temperaturetemperature
Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize
d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost
errdquo injectionerrdquo injection
A Live (attenuated) A Live (attenuated) vaccinesvaccines
B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate
responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe
toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves
artificial passive artificial passive immunization immunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
vaccinesvaccines Capsular polysaccharidesCapsular polysaccharides Inactivated protein exotoxins (toxoids)Inactivated protein exotoxins (toxoids) killed bacteriakilled bacteria Live attenuated BLive attenuated B Subcellular fragmentsSubcellular fragments Genes for Ags in some vectorsGenes for Ags in some vectors DNADNA
A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow
th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni
citycity are repeatedly selected are repeatedly selected BCGBCG
The mutation is expressed only The mutation is expressed only
under certain conditionsunder certain conditions
may be may be useful in vaccinesuseful in vaccines
Conditional-lethal mutationsConditional-lethal mutations
temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation
the organisms can the organisms can replicate replicate at a at a
relativelyrelatively low low permissive temperature permissive temperature
but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive
temperaturetemperature
Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize
d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost
errdquo injectionerrdquo injection
A Live (attenuated) A Live (attenuated) vaccinesvaccines
B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate
responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe
toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves
artificial passive artificial passive immunization immunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
A Live (attenuated) A Live (attenuated) vaccinesvaccines Consist of organisms attenuated by growConsist of organisms attenuated by grow
th in th in unfavourable conditionsunfavourable conditions The genes of organisms The genes of organisms mutatemutate Mutants Mutants lost virulence but retain antigenilost virulence but retain antigeni
citycity are repeatedly selected are repeatedly selected BCGBCG
The mutation is expressed only The mutation is expressed only
under certain conditionsunder certain conditions
may be may be useful in vaccinesuseful in vaccines
Conditional-lethal mutationsConditional-lethal mutations
temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation
the organisms can the organisms can replicate replicate at a at a
relativelyrelatively low low permissive temperature permissive temperature
but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive
temperaturetemperature
Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize
d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost
errdquo injectionerrdquo injection
A Live (attenuated) A Live (attenuated) vaccinesvaccines
B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate
responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe
toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves
artificial passive artificial passive immunization immunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
The mutation is expressed only The mutation is expressed only
under certain conditionsunder certain conditions
may be may be useful in vaccinesuseful in vaccines
Conditional-lethal mutationsConditional-lethal mutations
temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation
the organisms can the organisms can replicate replicate at a at a
relativelyrelatively low low permissive temperature permissive temperature
but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive
temperaturetemperature
Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize
d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost
errdquo injectionerrdquo injection
A Live (attenuated) A Live (attenuated) vaccinesvaccines
B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate
responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe
toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves
artificial passive artificial passive immunization immunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
temperature-sensitive conditional-temperature-sensitive conditional-lethal mutationlethal mutation
the organisms can the organisms can replicate replicate at a at a
relativelyrelatively low low permissive temperature permissive temperature
but but canrsquot grow at a highercanrsquot grow at a higher restrictive restrictive
temperaturetemperature
Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize
d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost
errdquo injectionerrdquo injection
A Live (attenuated) A Live (attenuated) vaccinesvaccines
B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate
responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe
toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves
artificial passive artificial passive immunization immunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
Can regain virulence by Can regain virulence by backmutationbackmutation Often induce stronger and better localizeOften induce stronger and better localize
d immunityd immunity Induce more appropriate responseInduce more appropriate response Do notDo not often often require adjuvantsrequire adjuvants or ldquoboost or ldquoboost
errdquo injectionerrdquo injection
A Live (attenuated) A Live (attenuated) vaccinesvaccines
B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate
responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe
toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves
artificial passive artificial passive immunization immunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
B Inactivated (dead) B Inactivated (dead) vaccinesvaccines Killed organisms or B productsKilled organisms or B products Induce Induce weak weak andor inappropriate andor inappropriate
responseresponse Immune memoryImmune memory may be variable or may be variable or poorpoor Usually Usually safesafe
toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves
artificial passive artificial passive immunization immunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
toxoidstoxoids Formaldehyde-treated exotoxinFormaldehyde-treated exotoxin Induce antitoxinInduce antitoxin Abs Abs No immunityNo immunity against the B themselves against the B themselves
artificial passive artificial passive immunization immunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
artificial passive artificial passive immunization immunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
Artificial passive Artificial passive immunizationimmunization Administer either preformed immunoreacAdminister either preformed immunoreac
tive tive serum or cellsserum or cells IgG is IgG is predominantpredominant ObtainedObtained from from human or animal donorshuman or animal donors
who have recovered from an infectious diwho have recovered from an infectious disease or been immunizedsease or been immunized
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization
Provide Provide immediate protectionimmediate protection Be useful for individuals Be useful for individuals
Cannot form AbsCannot form Abs Before Ab production Before Ab production
animal sera animal sera have the risk of allergichave the risk of allergic reac reactionstions
Artificial passive Artificial passive immunizationimmunization