16273B - en - 2012/07 ETEST APPLICATION GUIDE AEROBES RECOMMENDED INOCULUM INCUBATION SPECIFIC QUALITY CONTROL ORGANISM MEDIA 6) SUGGESTED MIC PANEL 3) COMMENTS 4) PHENOTYPE Suspension Temperature/ McF equivalent Time (h) Strain ATCC® medium Atmosphere Staphylococcus 29213TM ≤ 4 mg/L 35 ± 2°C MHA 0.5 Saline Cefoxitin (FX) 18-24h aureus Ambient air S. aureus 43300TM > 4 mg/L - Phenotype interpretation: MRSA OX ≥ 4 µg/mL and Mueller Hinton agar Oxacillin (OX) 35 ± 2°C TM XL ≥ 8 µg/mL 0.5-1 Saline 24h S. aureus 43300 (MHA) + 2% NaCl Amoxicillin/clavulanic acid (XL) Ambient air - Heavier inoculum improves detection of low level R - Phenotype interpretation: Oxacillin (OX) 35 ± 2°C BORSA MHA + 2% NaCl 0.5-1 Saline 24h S. aureus 29213TM OX ≥ 4 µg/mL and Amoxicillin/clavulanic acid (XL) Ambient air XL ≤ 4 µg/mL - Phenotype interpretation: OX ≥ 0.5 µg/mL Mueller Hinton agar 35 ± 2°C MRCNS 0.5-1 Saline Oxacillin (OX) 48h S. aureus 43300TM - Heavier inoculum (MHA) + 2% NaCl Ambient air improves detection of low Staphylococci level R - 0.1 mL inoculum/ 90mm plate - Phenotype interpretation: S. aureus VA ≥ 8µg/mL and 1) 24h. VISA/hVISA Brain Heart Infusion Vancomycin (VA) 35 ± 2°C QC ranges: TM TP ≥ 8µg/mL, 2 BHI broth Confirm at 29213 (macromethod) agar (BHI) 1) Teicoplanin (TP) 1) Ambient air TP = 0.5-2 µg/mL OR 48h VA = 1-4 µg/mL TP ≥12 µg/mL (alone) = VISA/hVISA - Confirm VISA/hVISA by PAP 18-24h. S. aureus 29213TM Etest GRD2) (Vancomycin (VA) / 35 ± 2°C - Confirm VISA/hVISA by VISA/hVISA MHA + 5% blood 0.5 BHI broth Confirm at S. aureus 700698TM Teicoplanin (TP) 1)) Ambient air PAP 48h S. aureus 700699TM - Phenotype interpretation: VISA/VRSA 35 ± 2°C VA ≥ 4µg/mL = VISA (standard MHA 0.5 Saline Vancomycin (VA) 24h S. aureus 29213TM Ambient air method) VA ≥ 16µg/mL = VRSA (CLSI) bioMérieux SA English - 1 Etest® Application Guide 16273B - en - 2012/07 RECOMMENDED INOCULUM INCUBATION SPECIFIC QUALITY CONTROL ORGANISM MEDIA 6) SUGGESTED MIC PANEL 3) COMMENTS 4) PHENOTYPE Suspension Temperature/ McF equivalent Time (h) Strain ATCC® medium Atmosphere Clindamycin (CM) Daptomycin (DPC) MRSA Linezolid (LZ) 16-20h. 35 ± 2°C Secondary MHA 0.5 Saline Vancomycin (VA) VA and TP S. aureus 29213TM Ambient air panel Teicoplanin (TP) 1) full 24h Quinupristin/dalfopristin (QDA) Rifampicin (RI) 1) Staphylococci Benzylpenicillin (PG) Erythromycin (EM) Non-MRSA Linezolid (LZ) 16-20h. 35 ± 2°C secondary MHA 0.5 Saline Trimethoprim/sulfamethoxazole (TS) VA and TP S. aureus 29213TM Ambient air panel Vancomycin (VA) full 24h Teicoplanin (TP) 1) Ciprofloxacin (CI) Ampicillin (AM) Daptomycin (DPC) 16-20h. Linezolid (LZ) Confirm VA 35 ± 2°C MHA 0.5 Saline Quinupristin/dalfopristin (QDA) E. faecalis 29212TM and TP at Ambient air Vancomycin (VA) 24h Teicoplanin (TP) 1) Minocycline (MC) Enterococci - Heavier inoculum preferable E. faecalis - SM > 1024 µg/mL or 24h. QC ranges: GM > 512 µg/mL = Gentamicin (GM) high range 35 ± 2°C HLAR MHA 0.5-1 Saline Confirm SM GM = 4-16 µg/mL 29212TM positive HLAR (CLSI) Streptomycin (SM) high range 1) Ambient air at 48h SM = 64–256 µg/mL - SM > 512 µg/mL or GM > 128 µg/mL = positive HLAR (EUCAST) Gentamicin (GM) Piperacillin/ tazobactam (PTc) 0.5 Cefepime (PM) 35 ± 2°C Escherichia coli 25922TM MHA (1 for mucoid Saline 16-20h Ciprofloxacin (CI) Ambient air E. coli 35218TM organisms) Imipenem (IP) (PTc) Aztreonam (AT) Enterobacteriaceae Cefotaxime/cefotaxime + clavulanic E. coli 35218TM acid (CT/CTL) Klebsiella 700603TM 0.5 Ceftazidime/ceftazidime + clavulanic 35 ± 2°C pneumoniae - Investigate ND results by ESBL MHA (1 for mucoid Saline 16-20h acid (TZ/TZL) Ambient air (ESBL positive) an alternative method organisms) Cefepime/cefepime + clavulanic acid Pseudomonas 27853TM (PM/PML) 1) aeruginosa bioMérieux SA English - 2 Etest® Application Guide 16273B - en - 2012/07 RECOMMENDED INOCULUM INCUBATION SPECIFIC QUALITY CONTROL ORGANISM MEDIA 6) SUGGESTED MIC PANEL 3) COMMENTS 4) PHENOTYPE Suspension Temperature/ McF equivalent Time (h) Strain ATCC® medium Atmosphere 0.5 MBL MP/MPI Meropenem/Meropenem +EDTA 35 ± 2°C K. pneumoniae 700603TM - Investigate ND results by MHA (1 for mucoid Saline 16-20h. 1) (MP/MPI) Ambient air K. pneumoniae BAA-2146TM an alternative method organisms) Enterobacteriaceae Klebsiella 700603TM pneumoniae Cefotetan/Cefotetan + Cloxacillin 35 ± 2°C - Investigate ND results by AmpC 2) MHA 0.5 Saline 16-20h (negative) (CN/CNI) Ambient air an alternative method K. pneumoniae BAA-1144TM (positive) Ceftazidime (TZ) Gentamicin (GM) 0.5 16-20h. Aztreonam (AT) 35 ± 2°C P. aeruginosa 27853TM MHA (1 for mucoid Saline 48h for slow Ciprofloxacin (CI) Ambient air E. coli 35218TM organisms) growers Imipenem (IP) (PTc) Pseudomonas spp. Piperacillin/tazobactam (PTc) P. aeruginosa 27853TM 0.5 16-20h. 35 ± 2°C (negative) - Investigate ND results by MBL IP/IPI 1) MHA (1 for mucoid Saline Imipenem/imipenem + EDTA (IP/IPI) 48h for slow Ambient air Stenotrophomonas 13636TM an alternative method organisms) growers maltophilia (positive) Ceftazidime (TZ) Meropenem (MP) 0.5 20-24h. Amikacin (AK) 35 ± 2°C P. aeruginosa 27853TM MHA (1 for mucoid Saline 48h for slow Ampicillin/sulbactam (AB) Ambient air E. coli 35218TM (AB) organisms) growers Levofloxacin (LE) Acinetobacter spp. Minocycline (MC) P. aeruginosa 27853TM 0.5 16-20h. 35 ± 2°C (negative) - Investigate ND results by MBL IP/IPI 1) MHA (1 for mucoid Saline Imipenem/imipenem + EDTA (IP/IPI) 48h for slow Ambient air Stenotrophomonas 13636TM an alternative method organisms) growers maltophilia (positive) Trimethoprim/sulfamethoxazole (TS) 0.5 Ceftazidime (TZ) 20-24h. 35 ± 2°C Burkholderia cepacia MHA (1 for mucoid Saline Levofloxacin (LE) 48h for slow E. coli 25922TM Ambient air organisms) Meropenem (MP) growers Minocycline (MC) Trimethoprim/sulfamethoxazole (TS) 0.5 Ceftazidime (TZ) 20-24h. Stenotrophomonas 35 ± 2°C E. coli 25922TM MHA (1 for mucoid Saline Levofloxacin (LE) 48h for slow maltophilia Ambient air E. coli 35218TM (TLc) organisms) Minocycline (MC) growers Ticarcillin/clavulanic acid (TLc) bioMérieux SA English - 3 Etest® Application Guide 16273B - en - 2012/07 FASTIDIOUS ORGANISMS RECOMMENDED INOCULUM INCUBATION SPECIFIC QUALITY CONTROL ORGANISM MEDIA 6) SUGGESTED MIC PANEL 3) COMMENTS 4) PHENOTYPE Suspension Temperature/ McF equivalent Time (h) Strain ATCC® medium Atmosphere Meropenem (MP) Cefotaxime (CT) MHA + 5% blood 0.5 MH broth or Benzylpenicillin (PG) 35 ± 2°C Streptococcus Pneumococci (CLSI) (1 for mucoid 20-24h 49619TM BHI broth Clindamycin (CM) 1) 5% CO pneumoniae MHF (EUCAST) organisms) 2 Vancomycin (VA) Trimethoprim/sulfamethoxazole (TS) Benzylpenicillin (PG) Cefotaxime (CT) MHA + 5% blood 0.5 Chloramphenicol (CL) MH broth or 35 ± 2°C Streptococci (CLSI) (1 for mucoid Ofloxacin (OF) 20-24h S. pneumoniae 49619TM BHI broth 5% CO MHF (EUCAST) organisms) Linezolid (LZ) 1) 2 Vancomycin (VA) Daptomycin (DPC) MH chocolate agar + Broth Abiotrophia & 35 ± 2°C 0.001% pyridoxal HCl + 1 (MH broth or 20-24h 5) 5) Granulicatella spp. 1) 5% CO 0.01% cysteine BHI broth) 2 Amoxicillin/ clavulanic acid (XL) Haemophilus Test 0.5 MH broth or Cefotaxime (CT) Haemophilus 49247TM Haemophilus 35 ± 2°C Media (HTM) (CLSI) (1 for mucoid HTM broth Meropenem (MP) 20-24h influenzae influenzae 5% CO MHF (EUCAST) organisms) (or BHI broth) Trimethoprim/sulfamethoxazole (TS) 2 H. influenzae 49766TM Chloramphenicol (CL) (MP, XM) MHA + 5% blood MH broth or 35 ± 2°C Moraxella catarrhalis 1) (CLSI) 0.5 20-24h 5) 5) BHI broth 5% CO MHF (EUCAST) 2 - For obligate anaerobes, ensure anaerobic Metronidazole (MZ) Brucella agar + 24-72h conditions are Brucella broth Clindamycin (CM) 5% blood + (48h for CM). 35 ± 2°C Bacteroides fragilis 25285TM maintained throughout or MH broth Cefoxitin (FX) Anaerobes vitamin K (1 µg/mL) + 1 Confirm all Anaerobic B. thetaiotaomicron 29741TM processing (or Schaedler Imipenem (IP) hemin (5 µg/mL) (BBA) S results at system Eubacterium lentum 43055TM - DO NOT VORTEX Broth + vit K3) Piperacillin/tazobactam (PTc) (CLSI) 48h. - Anaerobiosis must be Benzylpenicillin (PG) achieved within 1-2 hours for MZ bioMérieux SA English - 4 Etest® Application Guide 16273B - en - 2012/07 RECOMMENDED INOCULUM INCUBATION SPECIFIC QUALITY CONTROL ORGANISM MEDIA 6) SUGGESTED MIC PANEL 3) COMMENTS 4) PHENOTYPE Suspension Temperature/ McF equivalent Time (h) Strain ATCC® medium Atmosphere - 1 strip/90mm plate - Strip handle to be 35 ± 2°C placed at edge of plate. MH broth + Amoxicillin (AC) Micro- - H. pylori colonies are MHA + 5% blood 5% serum Clarithromycin (CH) aerophilic, Helicobacter pylori 1) 3 72h or longer Helicobacter pylori 43504TM pin-point, translucent (≥ 2 weeks old) (CLSI) or BHI broth + Metronidazole (MZ) except MZ - and difficult to see. 5% serum Tetracycline (TC) first 24h Tilt plate and use anaerobic oblique light to read endpoint. Ciprofloxacin (CI) Supplemented GC agar Benzylpenicillin (PG) MH broth 35 ± 2°C Neisseria - 3-4 strips max./150mm Gonococci (CLSI) 0.5 Tetracycline (TC) 20-24h 49226TM (or BHI broth) 5% CO gonorrhoeae plate to facilitate reading MH chocolate agar Ceftriaxone (TX) 2 Spectinomycin (SC) 1) Ciprofloxacin (CI) MHA + 5% blood Benzylpenicillin (PG) Broth 35 ± 2°C S. pneumoniae 49619TM Meningococci 1) (CLSI) 0.5 Trimethoprim/sulfamethoxazole (TS) 24h - Work in BSC (BHI broth) 5% CO E. coli 25922TM MH chocolate agar Meropenem (MP) 2 Ceftriaxone (TX) - 3-4 strips max./150mm plate to facilitate reading - Do not invert plate Ciprofloxacin (CI) MHA + 5% blood 35 ± 2°C - Campylobacter colonies Broth Gentamicin (GM) Campylobacter Campylobacter spp. 1) (CLSI) 1 48-72h