Haemophilus influenzae and Haemophilus parainfluenzae

TESTING CONDITIONS

• Medium: Disk diffusion: Haemophilus Test

Medium (HTM) Broth dilution: HTM broth

Haemophilus Test Medium

hematin stock solution is prepared by dissolving 50 mg of hematin powder in 100 mL of 0.01 mol/L NaOH with heat and stirring until the powder is thoroughly dissolved. 30 mL of the hematin stock solution and 5 g of yeast extract is added to 1 L of Mueller-Hinton agar (MHA) and autoclaved.After autoclaving and cooling, add 3 mL of a nicotinamide adenine dinucleotide (NAD) stock solution (50 mg of NAD dissolved in 10 mL of distilled water, filter sterilized) aseptically

Testing conditions

• Inoculum: Direct colony suspension, equivalent to a 0.5 McFarland standard

• Incubation: 35 ± 2 °C; • Disk diffusion: 5% CO2; 16 to 18 hours • Broth dilution: ambient air; 20 to 24 hours

Suggested grouping of antimicrobial agents that should be considered for routine testing and reporting for Haemophilus

spp.

• GROUP A DRUGS PRIMARILY TESTED AND REPORTED

AmpiciilinTrimethoprim-

Sulfamethoxazole

• GROUP B DRUGS PRIMARILY TESTED AND REPORTED SELECTIVELY

Ampicillin-SulbactamCefuroxime(parenteral)Cefotaxime or

ceftazidime or Ceftriaxone

ChloramphenicolMeropenem

Suggested grouping of antimicrobial agents that should be considered for routine testing and reporting for Haemophilus spp.

• GROUP C (SUPPLEMENTAL) - REPORT SELECTIVELY Azithromycin Clarithromycin Aztreonam Amoxycillin-Clavulanic acid Cefaclor Cefprozil Cefdinir or Cefixime orCefpodoxime Cefuroxime(oral) Ciprofloxacin or Levofloxacin or Lomefloxacin or Moxifloxacin or Ofloxacin Gemifloxacin Ertapenem or Imipenem Rifampin Telithromycin Tetracycline

MINIMAL QC Recommendations

Haemophilus influenzae ATCC® 49247

Haemophilus influenzae ATCC® 49766

Escherichia coli ATCC® 35218 (when testing

amoxicillin-clavulanic acid)

Amoxicillin-clavulanic acid, Azithromycin, Clarithromycin, Cefpodoxime,Cefixime, Cefuroxime,Cefaclor, Cefprozil, Loracarbef, Cefdinir, Telithromycin

Empiric therapy for respiratory tract infections

H. influenzae from CSF following drugs should be reported routinely.

Ampicillin

Third Generation Cephalosporins

Chloramphenicol

Meropenem

• The results of ampicillin susceptibility tests should be used to predict the activity of Amoxicillin.

• The majority of isolates of H.influenzae that are resistant to Ampicillin and Amoxicillin produce a TEM-type β-lactamase.

• In most cases, a direct β-lactamase test can provide a rapid means of detecting resistance to ampicillin and amoxicillin.

• Rare BLNAR strains of H. influenzae should be considered resistant to :-

Ampicillin-SulbactamAmoxacillin-Clavulanic acidCefuroximepiperacillin-tazobactamCefaclorCefetametCefonicid cefprozil Loracarbef

• Chloramphenicol not routinely reported on isolates of urinary tract.

• Rifampicin should not be used alone for antimicrobial therapy

Drug Conc. Of Drug used µm

H.Influenzae

Enterobacteriacae

Ampicillin 10 22 17

Ampillin-Sulbactam 10/10 20 15

Amoxicillin-Clavulanic acid

20/10 20 18

Piperacillin-Tazobactam

100/10 21 21

Cefotaxime or Ceftazidime or Ceftriaxone

303030

262626

262123

Cefuroxime 30 20 18

Cefpodoxime 10 21 21

Drug Conc. Of Drug used µm

H.influenzae Enterobacteriacae

Cefepime 30 26 18

Ciprofloxacin or Levofloxacn or Ofloxacin

555

181618

211716

Trimethoprim-Sulfamethoxazole

1.25/23.75 16 16

Chloramphenicol30

29 18

Meropenem 10 20 23

Drug Conc. Of Drug used µm

H.Influenzae

Enterobacteriacae

Tetracycline 30 29 15

Imipenem orEtrapenem

1010

1619

2323

Azithromycin orClarithromycin

1515

1213

_ _

Gatifloxacin 5 18 15

Rifampin 5 20 _

NEISSERIA GONORRHOEAE

TESTING CONDITIONS

• Medium: Disk Diffusion: GC agar base and 1% defined

growth supplement

Agar dilution: GC agar base and 1% defined growth

supplement

GC agar supplement 1.1g L-cysteine 0.03g guanine HCL 3 mg thiamine HCL 13 mg para-aminobenzoic acid[PABA] 0.1 g B12 0.1 g cocarboxylase 0.25 g NAD 1 g adenine 10 g L-glutamine 100 g glucose 0.02 g ferric nitrate[in 1 L H2O]

• Inoculum:Direct colony suspension, equivalent to a 0.5 McFarland standard

• Incubation: 36±1 0c, 5% CO 2,

20 to 24 hours

TESTING CONDITIONS

MINIMAL QC Recommendations

Neisseria gonorrhoeae ATCC 49226

Suggested grouping of antimicrobial agents that should be considered for routine testing and reporting for Neisseria

gonorrhoeae

• GROUP C (SUPPLEMENTAL)-REPORT SELECTIVELYCefixime or CefpodoximeCefotaxime or ceftriaxoneCefoxitinCefuroximeCiprofloxacin or OfloxacinPenicillinSpectinomycinTetracycline

single dose treatment regimen for Neisseria gonorrhoeae

Ceftriaxone (125 mg IM)

Cefixime (400mg PO)

Ciprofloxacin (500 mg PO)

Ofloxacin (400 mg PO)

Levofloxacin (250 mg PO)

Drug Conc. Of drug µm Zone

Penicillin 10 units 47

Cefotaxime or Ceftriaxone

3030

3135

Cefoxitin 30 28

Cefepime 30 31

Ceftazidime 30 31

Cefpodoxime 10 29

Tetracycline 30 38

Ciprofloxacin or ofloxacin

55

4131

Standard Zone diameters for AST of Neisseria gonorrhoeae