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05/03/2023 Dr.T.V.Rao MD 1
BLOOD CULTURINGusing automation
Dr.T.V.Rao MD
05/03/2023 Dr.T.V.Rao MD 2
Need for Blood Culture?
No microbiological test is more essential to the
clinician than the blood culture. The finding of
pathogenic microorganisms in a patient’s
bloodstream is of great importance in terms of
diagnosis, prognosis, and therapy.” - L. Barth Reller, Clin. Infect. Diseases, 1996
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Sources of Bacteremia Spread
•Pericarditis and Peritonitis•Pneumonias•Pressure sores•Prosthetic medical devices•Total hip replacement•Skeletal system•Skin and soft tissue
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Blood culturing most important and life
saving InvestigationNeeds optimal Methods for Diagnosis
of Blood Borne Pathogens
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Blood Collection•Aseptic collection procedure is critical Amount of blood•1:10 ratio of blood to broth•Younger than 10 years – 1 ml of blood for every year of life•Over 10 years – 20 ml
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Blood Collection •Frequency of Collection•Depends if bacteremia is
transient, intermediate or continuous•Number of cultures
collected are usually inversely related to the type of bacteremia•Usually x3 from different
body sites
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Venipuncture• Venipuncture is the process of obtaining intravenous access for the purpose of intravenous therapy or obtaining a sample of venous blood. This procedure is performed by medical laboratory scientists, medical practitioners, some EMTs, paramedics phlebotomists and other nursing staff. Venipuncture is one of the most routinely performed invasive procedures and is carried out for two reasons, to obtain blood for diagnostic purposes or to monitor levels of blood components (Lavery & Ingram 2005).
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LABELING THE SAMPLE
•Properly labelled sample is essential so that the results of the test match the patient. The key elements in labelling are:•Patient's surname, first and middle.•Patient's ID number.•NOTE: Both of the above MUST match the
same on the requisition form.•Date, time and initials of the phlebotomist
must be on the label of EACH tube.
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Principles for Collection
•Gloves will be worn in accordance with standard precautions.••Appropriate verification of the patient's identity, by
means of an armband or area specific procedure, will occur before the specimen collection.••Cultures should be drawn before administration of
antibiotics, if possible. ???•• blood cultures should be drawn from lines, but
should be drawn viavenipuncture.
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The requisitions form should be completely filled out, and the
requisition must indicate the tests ordered.
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Self Protection
A few ways to make sure your role in the collection process is carried out with efficiency, orderliness and
safety
Steps 1 – 3, Check, Explain, Wash
•1.Identify the patient •2.Explain the procedure to the patient.•3.Wash hands with soap and water with friction for 15 seconds or use alcohol based hand rub
Dr.T.V.Rao MD 1305/03/2023
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Materials• Chlorhexidine swabs (1-2 packages)• Alcohol swabs• Blood culture bottles (2 bottles per set)• 2 syringes (adult: 20 cc, paediatric: 5 cc)• 2 needles (adult: 22 gauge or preferably larger butterfly or
standard needle; pediatric: 25 or 23 gauge butterfly or standard needle)• Gloves (sterile &nonsterile)• Tourniquet• Sterile gauze pad• Adhesive strip or tape• Self-sticking patient labels• Plastic zip lock specimen bags
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. Barrier protection for the phlebotomist consists of the latex
gloves.
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Obtaining Blood • Locate the vein• Prep kit• Alcohol 5 sec. Dry 30-60 sec ( resource poor conditions )• Ideal to collect with alcohol swabs containing 2% Chlorhexidine and
70% isopropyl alcohol• Remove caps, clean with alcohol• Put on gloves• Without palpating, draw 20 ml and put 10 in anaerobic and 10 in
aerobic bottle• Dispose of syringe in sharps container• Label bottles and send to lab
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Method of Blood Collection
• A minimum of 10 ml of blood is taken through venipuncture and injected into two or more "blood bottles" with specific media for aerobic and anaerobic organisms.• The blood is collected using clean
technique. This requires that both the tops of the culture bottles and the venipuncture site of the patient are cleaned prior to collection with alcohol swabs containing 2% Chlorhexidine and 70% isopropyl alcohol.
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The area of skin is cleaned with a disinfectant, or an alcohol swab.
• Using sterile gloves, do not wipe away the surgical solution, touch the puncture site, or in any way compromise the sterile process. It is vital that the procedure is performed in as sterile a manner as possible as the persistent presence of skin commensals in blood cultures could indicate endocarditis but they are most often found as contaminants
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The vein is anchored and the needle is
inserted.
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The vacutainer tube is depressed into the needle to begin drawing
blood
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Additional vacutainer tubes can be utilized. Determine what tests are ordered and what tubes will be necessary BEFORE you begin to
draw blood, and determine the order of draw for the tubes. .
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When the final tube is being drawn, release the tourniquet. Then remove the tube, and remove the needle.
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After the needle is removed from the vein, apply firm pressure over the site to achieve haemostasis.
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Apply a bandage to the area.
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Preparation of Cap before Injecting Blood
•Prep the rubber cap of the blood culture bottles with an alcohol pad in a circular motion. Allow the alcohol to dry.
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Inject the Blood …..•Inject the blood into the Selected Media•Gently rotate the bottles to mix the blood & the broth (do not shake vigorously).
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Follow the universal precautions when disposing Needle
•Dispose of needle in sharps container and dispose of other waste in proper container
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Label the tubes, checking the requisition for the proper
identification.
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Give the all possible Medical Information
•Patient’s name•• Hospital number (Patient ID)•• Patient’s location (room and bed #)•• Date and time of collection•• Collector’s initials•• Site of venipuncture•• Or other information as per facility• Include you Mobile Contact No – A vital
information can be delivered any time
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Document the Medical Records
•Document the following in the medical record:•–Date & time specimen obtained•–Site of specimen collection
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Frequency of Collection
•Frequency of Collection•Depends if bacteremia is transient, intermediate or continuous•Number of cultures collected are usually inversely related to the type of bacteremia•Usually x3 from different body sites
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Second Set
•If 2 or more sets of blood cultures have been ordered, obtain the second set in the same manner as the first, making a new venipuncture at a different site.
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Bacteria and Fungi Are Identified by Phenotypic Characters
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Biochemical Tests gives Better Clues in Identification
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Newer Blood Culture Methods
•Newer Blood Culture Systems•Biphasic Broth-Slide System• Agar “paddles” attached to top of bottle• Closed system
•Continuous Monitoring Blood Culture Systems• BacTec – measures 14CO2• BacTec 9000 Series – measures CO2• ESP – measures consumption of gases• BacT-Alert – measures change in pH
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Newer Technologies available at our Hospital •Over the past few years, dramatic improvement has taken place in blood culture methods, media, and systems. Most of the technologically advanced blood culture systems are fully automated continuously monitored blood culture systems. These systems electronically monitor blood culture bottles every 8-10 minutes and detect algorithms based on assessments of changes associated with microbial growth.
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SEPTIC SHOCK A PRIORITY
• Let us think of septic shock as a top priority, at least we receive 10% of the samples, we process are Blood cultures and the number of specimens increase with the expansion of ICU and MICU care, According to the literature, the risk of death from septic shock increases by over 7% with every hour that passes from the onset of shock until the start of targeted therapy. tools in clinical microbiology are primarily based on techniques that evolved 30 to 40 years ago
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Reliable Microorganism Recovery
• BacT/ALERT® Culture Media provides a wide range of media bottles to rapidly and reliably recover an array of microorganisms, including bacteria, mycobacteria, fungi, and yeasts from various sample types. BacT/ALERT® Culture Media is FDA-cleared for blood, sterile body fluids, and platelets, and can detect 98% of isolates within 72 hours.* -
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Automation reduces the time requirement
• But this can be completed
within 30 hours by using automated techniques. This is especially useful when large number of specimens needs to be cultured, as the instrument, which has been programmed for the same, automatically screens these.
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BacT/AlerT 3D culture system
BacT/AlerT 3D culture system. This is the first automated non-radiometric and non-invasive culture system that continuously monitors system for culture of bacteria (both aerobic and anaerobic), fungi and mycobacteria. All these bacteria can be cultured using different media as prescribed..
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bioMérieux BacT/ALERT® 3D The bioMérieux
BacT/ALERT® 3D provides an optimal environment for the recovery of a wide range of pathological organisms, including bacteria, yeasts and mycobacteria; utilizing proprietary plastic culture bottles ensuring added safety to the user.
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BacT/ALERT® 3D Microbial Detection System
• This newest generation of the time-tested BacT/ALERT system offers advantages in every dimension of testing. From its space-saving modular design to its easy touch-screen operation and flexible data management options, every laboratory will find something to love about the BacT/ALERT 3D!
Principles of functioning of BacT alert Monitors
Microorganisms multiply in the media, generating CO2. As CO2 increases, the sensor in the bottle turns a lighter colour.
Measuring reflected light, the BacT/ALERT 3D monitors and detects color changes in the sensor.
Algorithms analyze the data to determine positivity, and the laboratory is notified immediately with visual and audible alarms.
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Principles in BacT/AlerT 3D culture system
This is a closed system and works on the colorimetric principle of detection of CO2 produced by the organisms. The CO2 causes a lowering of the pH of the medium, which in turn produces a colour change in a sensor attached to the CO2-sensitive base of each bottle.
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What are other Specimens sent for culturing
•Cerebrospinal fluid•Pleural Aspirates •Joint Aspirates•Peritoneal fluid •CT guided Aspirates
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Specimens Not suitable for BacT/AlerT 3D culture system
•Urine•Stool•Pus •Aspirates from Surgical
Site Infections•Any specimen
contaminated with contaminants and commensals including oral secretions
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CT guided Aspirates •CT guided aspirate is a valuable material we can try best out of the aspirates
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WHO IS ACCOUNTABLE TO RUNNING THE SYSTEM
• 1The physicians should give many clinical details why they wish to culture blood, as automation is fraught with many errors if the coordination fails between treating doctor and laboratory personal the errors with automation are more expensive dangerous with loss of valuable time and the purpose of automation is lost • 2 All the requests should be identified with caring Doctor or
nurse responsible for treatment decision and better with active Mobile contact number for faster communication in case of any growth as the machine alerts every 10 minutes a valuable time is saved for faster decisions
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WHO IS ACCOUNTABLE TO RUNNING THE SYSTEM
•3 The Medical Microbiologists should be well know ledged to identify the contamination and source to make the system running with scientific efficacy • 4 With experience I wish to say automation needs more knowledge of Medical Microbiologists or the matters will be counterproductive in false identification of contaminants
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WHO IS ACCOUNTABLE TO RUNNING THE SYSTEM
• 5 A regular audit of the functioning and reporting both by internal and external experts. • 6 I wish who wish to run the Automation must read from
more at Bailey and Scot diagnostic microbiology available in our libraries • 7 Ill qualified and careless Microbiologists, and technicians
are more dangerous as the errors at many stages multiplies faster than traditional methods of culturing leasing greater misuse of Antibiotics
Automation improves quality of services
Overall, laboratories transitioning from conventional to automated processes find that technologists and microbiologists are more open to innovation and improved quality.
Dr.T.V.Rao MD 5105/03/2023
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Automation Signals Bacteremia cases
•After inoculating the culture vials, they are sent to the clinical pathology microbiology department. Here the bottles are entered into a blood culture machine, which incubate the specimens at body temperature. The blood culture instrument reports positive blood cultures (cultures with bacteria present, thus indicating the patient is "bacteremia"). Most cultures are monitored for 5 days after which negative vials are removed.
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The positives cases to be proceeded without delay•A vial is positive, a microbiologist will perform a
Gram Stain on the blood for a rapid, general ID of the bacteria, which they will report to the attending physician of the bacteremic patient. The blood is also subcultured onto agar plates to isolate the pathogenic organism for culture and susceptibility testing, which takes up to 3 days. This culture & sensitivity (C&S) process identifies the species of bacteria. Antibiotic sensitivities are then assessed on the bacterial isolate to inform clinicians on appropriate antibiotics for treatment.
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Rapid Susceptibility Testing
•Rapid susceptibility will be carried out for gram negative and staphylococcal isolates and other isolates on request. These will be reported within 12 hours using API systems. Automation has made it easier to arrive at a precise laboratory diagnosis of infection
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VITEK® MS•VITEK® MS Mass spectrometry microbial
identification system An automated mass spectrometry microbial identification system that uses Matrix Assisted Laser Desorption Ionization Time-of-Flight (MALDI-TOF) technology and a comprehensive database of clinically relevant species for results in minutes Robust & accurate ID with Advanced Spectra Classifier Seamless integration of ID/AST results for optimized workflow Complete traceability & flexibility -
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Rapid & clear identification
•VITEK® MS is an innovative, automated microbial identification system that uses MALDI-TOF (Matrix Assisted Laser Desorption Ionization Time-of-Flight) technology. In just minutes, this mass spectrometry technology can provide clear identification at the species, genus and family level. You can provide clinicians with quick information to start appropriate treatment, contributing to overall patient care and outcomes.
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The Contaminated Blood Culture
•If the skin is not adequately cleansed before drawing blood for culture, bacteria on the skin will be injected into the bottle, producing a false positive blood culture•It is difficult for the physician to determine whether the bacteria growing in the blood culture is a real pathogen causing bloodstream infection or whether bacteria on the skin have contaminated the culture. This can lead to excess use of antibiotics and prolongation of hospital stay.
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•Program Created by Dr.T.V.Rao MD for Medical and Microbiology
Professionals Globally for improving the Skills in Diagnostic Microbiology
•Email•[email protected]
