European Committee on Antimicrobial Susceptibility Testing

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1 European Committee on Antimicrobial Susceptibility Testing Routine and extended internal quality control as recommended by EUCAST Version 5.0, valid from This document should be cited as "The European Committee on Antimicrobial Susceptibility Testing. Routine and extended internal quality control as recommended by EUCAST. Version 5.0, General Page Notes 1 Changes Routine quality control Page Escherichia coli ATCC 59 Escherichia coli ATCC Pseudomonas aeruginosa ATCC Staphylococcus aureus ATCC 91 7 Enterococcus faecalis ATCC 91 8 Streptococcus pneumoniae ATCC Haemophilus influenzae NCTC Haemophilus influenzae ATCC Campylobacter jejuni ATCC Extended quality control for detection of resistance mechanisms with disk diffusion Page Klebsiella pneumoniae ATCC Staphylococcus aureus NCTC Enterococcus faecalis ATCC Haemophilus influenzae ATCC

2 EUCAST QC Tables v. 5.0, valid from Notes 1. In EUCAST quality control (QC) tables, both ranges and targets are listed. Repeat testing of EUCAST quality control strains should yield individual and zone diameter values randomly distributed within the recommended ranges. If the number of tests is 10, the mode should be the target value and the mean zone diameter should be close to the target value.. For access to ISO standard documents, see EUCAST quality control strains for routine QC are used to monitor test performance. Control tests should be set up and checked daily, at least for antibiotic agents which are part of routine panels. For analysis of the QC test results, see EUCAST Disk Diffusion Manual, section E. coli ATCC 518 (TEM-1 β-lactamase-producing strain) is recommended to check the inhibitor component of penicillin inhibitor-combination disks for antimicrobial susceptibility testing of Enterobacteriaceae and should be part of the routine QC. E. coli ATCC 59 is used to check the active component. 5. EUCAST quality control strains for extended QC are complementary to the EUCAST routine quality control strains. These strains are recommended for detection of specific resistance mechanisms (ESBL, MRSA, VRE, HLGR and PBP mutations) and are used to check that routine susceptibility testing will result in the correct S, I and R categorisation. Extended QC should be performed with any change in the susceptibility testing system (with each new batch of disks or medium) and/or monthly. 1

3 EUCAST QC Tables v. 5.0, valid from Changes from previous version Version General ATCC 59 ATCC 518 ATCC NCTC 8468 ATCC Changes Cells containing a change or an addition from EUCAST Routine QC tables v. 4.0 or Extended QC tables v 1.0 are marked yellow. EUCAST QC strains for routine and extended quality control presented in one table. New recommendations for quality control of penicillin inhibitor-combination disks. Included in routine quality control of penicillin inhibitor-combination disks. QC ranges added: Amoxicillin-clavulanic acid, ampicillin-sulbactam and piperacillin-tazobactam (). Ticarcillin-clavulanic acid ( and zone diameter). Alternative test with S. aureus ATCC 91 added for oxacillin 1 µg. H. influenzae NCTC 8468 exhibits unusual growth characteristics and will be excluded from 016. QC ranges added: Amoxicillin range. Zone diameter ranges for amoxicillin-clavulanic acid, ampicillin, benzylpenicillin, cefepime, cefixime, cefotaxime, cefpodoxime, cefuroxime, chloramphenicol, doripenem, ertapenem, erythromycin, imipenem, meropenem, minocycline, nalidixic acid, rifampicin, telithromycin, tetracycline and trimethoprimsulfamethoxazole.

4 Routine QC EUCAST QC Tables v. 5.0, valid from Escherichia coli ATCC 59 (NCTC 141, CIP 76.4, DSM 110, CCUG 1760, CECT 44) Disk diffusion methodology: Mueller-Hinton agar, McFarland 0.5, air, 5±1ºC, 18±h. Read zone edges as the point showing no growth viewed from the back of the plate against a dark background illuminated with reflected light. Target 1 Range (µg) Target 1 Range Amikacin Amoxicillin Amoxicillin-clavulanic acid 4, Ampicillin Ampicillin-sulbactam 5, Aztreonam Cefadroxil Cefalexin Cefepime Cefixime Cefotaxime Cefoxitin Cefpodoxime Ceftaroline Ceftazidime Ceftibuten Ceftobiprole IP IP IP Ceftriaxone Cefuroxime Chloramphenicol Ciprofloxacin Colistin Doripenem Ertapenem Fosfomycin Gentamicin Imipenem Levofloxacin Mecillinam Meropenem Moxifloxacin Nalidixic acid Netilmicin Nitrofurantoin Norfloxacin Ofloxacin Pefloxacin Piperacillin Piperacillin-tazobactam 5, Ticarcillin Ticarcillin-clavulanic acid 4, Tigecycline Tobramycin Trimethoprim Trimethoprim-sulfamethoxazole

5 Routine QC EUCAST QC Tables v. 5.0, valid from Escherichia coli ATCC 59 (NCTC 141, CIP 76.4, DSM 110, CCUG 1760, CECT 44) From International Standards Organisation, ISO : 006 (with updates as in the latest CLSI M100 document), except ranges in bold/italics established by EUCAST. From Clinical and Laboratory Standards Institute, M100-S5, 015, except ranges in bold/italics established by EUCAST. All ranges have been validated by EUCAST. 4 For testing, the concentration of clavulanic acid is fixed at mg/l. 5 E. coli ATCC 518 (TEM-1 β-lactamase-producing strain) is used to check the inhibitor component. 6 Ignore growth that may appear as a thin inner zone on some batches of Mueller-Hinton agar. 7 For testing, the concentration of sulbactam is fixed at 4 mg/l. 8 For fosfomycin determination, the medium must be supplemented with glucose-6-phosphate to a final concentration of 5 mg/l. 9 For testing, the concentration of tazobactam is fixed at 4 mg/l. 10 For tigecycline broth microdilution determination, the medium must be prepared fresh on the day of use. 11 Trimethoprim:sulfamethoxazole in the ratio 1:19. values are expressed as the trimethoprim concentration. IP = In Preparation 4

6 Routine QC EUCAST QC Tables v. 5.0, valid from Escherichia coli ATCC 518 (NCTC 11954, CIP 10181, DSM 59, CCUG 0600, CECT 94) TEM-1 β-lactamase-producing strain (non-esbl) used to check the inhibitor component of penicillin inhibitpr combination disks. Disk diffusion methodology: Mueller-Hinton agar, McFarland 0.5, air, 5±1ºC, 18±h. Read zone edges as the point showing no growth viewed from the back of the plate against a dark background illuminated with reflected light. Target 1 Range (µg) Target 1 Range Amoxicillin-clavulanic acid Ampicillin-sulbactam Piperacillin-tazobactam Ticarcillin-clavulanic acid From Clinical and Laboratory Standards Institute, M100-S5, 015, except ranges in bold/italics established by EUCAST. All ranges have been validated by EUCAST. For testing, the concentration of clavulanic acid is fixed at mg/l. 4 Ignore growth that may appear as a thin inner zone on some batches of Mueller-Hinton agar. 5 For testing, the concentration of sulbactam is fixed at 4 mg/l. 6 For testing, the concentration of tazobactam is fixed at 4 mg/l. 5

7 Routine QC EUCAST QC Tables v. 5.0, valid from Pseudomonas aeruginosa ATCC 785 (NCTC 190, CIP , DSM 1117, CCUG 17619, CECT 108) Disk diffusion methodology: Mueller-Hinton agar, McFarland 0.5, air, 5±1ºC, 18±h. Read zone edges as the point showing no growth viewed from the back of the plate against a dark background illuminated with reflected light. Target 1 Range (µg) Target 1 Range Amikacin Aztreonam Cefepime Ceftazidime Ciprofloxacin Colistin Doripenem Fosfomycin Gentamicin Imipenem Levofloxacin Meropenem Netilmicin Piperacillin Piperacillin-tazobactam Ticarcillin Ticarcillin-clavulanic acid Tobramycin From International Standards Organisation, ISO : 006 (with updates as in the latest CLSI M100 document). From Clinical and Laboratory Standards Institute, M100-S5, 015, except ranges in bold/italics established by EUCAST. All ranges have been validated by EUCAST. 4 For fosfomycin determination, the medium must be supplemented with glucose-6-phosphate to a final concentration of 5 mg/l. 5 For testing, the concentration of tazobactam is fixed at 4 mg/l. 6 For testing, the concentration of clavulanic acid is fixed at mg/l. 6

8 Routine QC EUCAST QC Tables v. 5.0, valid from Staphylococcus aureus ATCC 91 (NCTC 197, CIP 1049, DSM 569, CCUG 15915, CECT 794) β-lactamase-producing strain (weak) Disk diffusion methodology: Mueller-Hinton agar, McFarland 0.5, air, 5±1ºC, 18±h. Read zone edges as the point showing no growth viewed from the back of the plate against a dark background illuminated with reflected light. Target 1 Range (µg) Target 1 Range Amikacin Ampicillin Azithromycin Benzylpenicillin unit Cefoxitin Ceftaroline Ceftobiprole IP IP IP Chloramphenicol Ciprofloxacin Clarithromycin Clindamycin Daptomycin Doxycycline Erythromycin Fosfomycin Fusidic acid Gentamicin Levofloxacin Linezolid Minocycline Moxifloxacin Mupirocin Netilmicin Nitrofurantoin Norfloxacin Ofloxacin Quinupristin-dalfopristin Rifampicin Teicoplanin Telavancin Tetracycline Tigecycline Tobramycin Trimethoprim Trimethoprim-sulfamethoxazole Vancomycin From International Standards Organisation, ISO : 006 (with updates as in the latest CLSI M100 document). Established and validated by EUCAST. 4 From Clinical and Laboratory Standards Institute, M100-S5, 015, and validated by EUCAST. 5 For daptomycin determination, the medium must be supplemented with Ca + to a final concentration of 50 mg/l. 6 For fosfomycin determination, the medium must be supplemented with glucose-6-phosphate to a final concentration of 5 mg/l. 7 For telavancin determination, the medium must be supplemented with polysorbate-80 to a final concentration of 0.00%. 8 For tigecycline broth microdilution determination, the medium must be prepared fresh on the day of use. 9 Trimethoprim:sulfamethoxazole in the ratio 1:19. values are expressed as the trimethoprim concentration. IP = In Preparation 7

9 Routine QC EUCAST QC Tables v. 5.0, valid from Enterococcus faecalis ATCC 91 (NCTC 1697, CIP 1014, DSM 570, CCUG 9997, CECT 795) Disk diffusion methodology: Mueller-Hinton agar, McFarland 0.5, air, 5±1ºC, 18±h. Read zone edges as the point showing no growth viewed from the back of the plate against a dark background illuminated with reflected light. Target 1 Range (µg) Target 1 Range Ampicillin Ciprofloxacin Gentamicin Imipenem Levofloxacin Linezolid Nitrofurantoin Norfloxacin Quinupristin-dalfopristin Teicoplanin Tigecycline Trimethoprim Trimethoprim-sulfamethoxazole Vancomycin From International Standards Organisation, ISO : 006 (with updates as in the latest CLSI M100 document). Established and validated by EUCAST. 4 Screening disk for high-level aminoglycoside-resistance in enterococci. 5 For tigecycline broth microdilution determination, the medium must be prepared fresh on the day of use. 6 From Clinical and Laboratory Standards Institute, M100-S5, Trimethoprim:sulfamethoxazole in the ratio 1:19. values are expressed as the trimethoprim concentration. 8

10 Routine QC EUCAST QC Tables v. 5.0, valid from Streptococcus pneumoniae ATCC 49619* (NCTC 1977, CIP 10440, DSM 11967, CCUG 68) Strain with reduced susceptibility to benzylpenicillin * Zone edges for S. pneumoniae on MH-F are often accompanied by α-haemolysis. Read inhibition of growth and not inhibition of haemolysis. Tilt the plate to easier differentiate between haemolysis and growth. There is usually growth in the whole area of α-haemolysis but on some MH-F media, there is additional α-haemolysis without growth. Disk diffusion methodology: Mueller-Hinton agar + 5% defibrinated horse blood and 0 mg/l β-nad, McFarland 0.5, 5% CO, 5±1ºC, 18±h. Read zone edges as the point showing no growth viewed from the front of the plate with the lid removed and with reflected light. Target 1 Range (µg) Target 1 Range Amoxicillin Ampicillin Azithromycin Benzylpenicillin unit Cefaclor Cefepime Cefotaxime Cefpodoxime Ceftaroline IP IP Ceftobiprole IP IP IP Ceftriaxone Cefuroxime Chloramphenicol Ciprofloxacin Clarithromycin Clindamycin Daptomycin Doripenem Doxycycline Ertapenem Erythromycin Imipenem Levofloxacin Linezolid Meropenem Minocycline Moxifloxacin Nitrofurantoin Norfloxacin Ofloxacin Oxacillin Rifampicin Teicoplanin Telithromycin Tetracycline Tigecycline Trimethoprim-sulfamethoxazole Vancomycin From International Standards Organisation, ISO : 006 (with updates as in the latest CLSI M100 document). Established and validated by EUCAST. 4 Clinical and Laboratory Standards Institute, M100-S5, 015, and validated by EUCAST. 5 For daptomycin determination, the medium must be supplemented with Ca + to a final concentration of 50 mg/l. 6 S. aureus ATCC 91 can be used for quality control of oxacillin 1 µg with target mm and range 19-5 mm. 7 For tigecycline broth microdilution determination, the medium must be prepared fresh on the day of use. 8 Trimethoprim:sulfamethoxazole in the ratio 1:19. values are expressed as the trimethoprim concentration. IP = In Preparation 9

11 Routine QC EUCAST QC Tables v. 5.0, valid from Haemophilus influenzae NCTC (CIP 54.94, CCUG 946) Disk diffusion methodology: Mueller-Hinton agar + 5% defibrinated horse blood and 0 mg/l β-nad, McFarland 0.5, 5% CO, 5±1ºC, 18±h. Read zone edges as the point showing no growth viewed from the front of the plate with the lid removed and with reflected light. Target Range (µg) Target Range Amoxicillin-clavulanic acid Note 1 Note Ampicillin Note 1 Note Benzylpenicillin Note 1 Note 1 1 unit Cefaclor Note 1 Note Cefepime Note 1 Note Cefixime Note 1 Note Cefotaxime Note 1 Note Cefpodoxime Note 1 Note Ceftaroline Note 1 Note Ceftibuten Note 1 Note Ceftriaxone Note 1 Note Cefuroxime Note 1 Note Chloramphenicol Note 1 Note Ciprofloxacin Note 1 Note Doripenem Note 1 Note Ertapenem Note 1 Note Erythromycin Note 1 Note Imipenem Note 1 Note Levofloxacin Note 1 Note Meropenem Note 1 Note Minocycline Note 1 Note Moxifloxacin Note 1 Note Nalidixic acid Note 1 Note Ofloxacin Note 1 Note Rifampicin Note 1 Note Telithromycin Note 1 Note Tetracycline Note 1 Note Trimethoprim-sulfamethoxazole Note 1 Note H. influenzae NCTC 8468 exhibits unusual growth characteristics and will be excluded from 016. Calculated by EUCAST. Established and validated by EUCAST. 10

12 Routine QC EUCAST QC Tables v. 5.0, valid from Haemophilus influenzae ATCC (NCTC 1975, CIP 10570, DSM 11970, CCUG 959) Disk diffusion methodology: Mueller-Hinton agar + 5% defibrinated horse blood and 0 mg/l β-nad, McFarland 0.5, 5% CO, 5±1ºC, 18±h. Read zone edges as the point showing no growth viewed from the front of the plate with the lid removed and with reflected light. Target 1 Range (µg) Target Range Amoxicillin-clavulanic acid Amoxicillin Ampicillin Ampicillin-sulbactam Azithromycin Benzylpenicillin unit Cefepime Cefixime Cefotaxime Cefpodoxime Ceftaroline IP IP Ceftibuten IP IP Ceftriaxone IP IP Cefuroxime Chloramphenicol Ciprofloxacin IP IP Clarithromycin Doripenem Doxycyline Ertapenem Erythromycin Imipenem Levofloxacin IP IP Meropenem Minocycline Moxifloxacin IP IP Nalidixic acid Ofloxacin IP IP Rifampicin Roxithromycin Telithromycin Tetracycline Trimethoprim-sulfamethoxazole Established and validated by EUCAST. For testing, the concentration of clavulanic acid is fixed at mg/l. 4 For testing, the concentration of sulbactam is fixed at 4 mg/l. 5 From Clinical and Laboratory Standards Institute, M100-S5, 015, and validated by EUCAST. 6 Trimethoprim:sulfamethoxazole in the ratio 1:19. values are expressed as the trimethoprim concentration. IP = In Preparation 11

13 Routine QC EUCAST QC Tables v. 5.0, valid from Campylobacter jejuni ATCC 560 (NCTC 1151, CIP 70, DSM 4688, CCUG 1184) Disk diffusion methodology: Mueller-Hinton agar + 5% defibrinated horse blood and 0 mg/l β-nad, McFarland 0.5, microaerobic environment, 41±1ºC, 4h. Read zone edges as the point showing no growth viewed from the front of the plate with the lid removed and with reflected light. The MH-F plates should be dried prior to inoculation to reduce swarming (at 0-5 C over night or at 5 C, with the lid removed, for 15 min). (µg) Target Range Target 1 Range Ciprofloxacin IP IP Erythromycin IP IP Tetracycline IP IP Established and validated by EUCAST. IP = In Preparation 1

14 Extended QC EUCAST QC Tables v. 5.0, valid from Quality control strains for detection of resistance mechanisms with disk diffusion on Mueller-Hinton agar Disk diffusion methodology: Mueller-Hinton agar, McFarland 0.5, air, 5±1ºC, 18±h. Read zone edges as the point showing no growth viewed from the back of the plate against a dark background illuminated with reflected light. Klebsiella pneumoniae ATCC (NCTC 168, CCUG 4541, CECT 7787) SHV-18 ESBL producer (µg) Target Range susceptibility 1 Aztreonam 0 R 9-17 Cefotaxime 5 I or R 1-18 Cefpodoxime 10 R 9-16 Ceftazidime 10 I or R 6-1 Ceftriaxone 0 I or R 16- Comments Staphylococcus aureus NCTC 149 Methicillin resistant (MRSA), meca positive (µg) Target Range susceptibility 1 Cefoxitin 0 R 14-0 Comments Enterococcus faecalis ATCC 5199 (NCTC 179,CIP , DSM 1956, CCUG 489) High-level gentamicin resistant (HLGR) (µg) Gentamicin 0 R 6 Teicoplanin 0 S 16-0 Vancomycin 5 R 6-1 Examine zone edge with transmitted light (plate held up to light). Inhibition zones with fuzzy zone edges are interpreted as resistant, even if the zone diameter is above the susceptible breakpoint. 1 Targets comply with EUCAST clinical breakpoints and are set to ensure that resistance mechanisms are correctly detected. Interpretation according to EUCAST clinical breakpoints: S=Susceptible, I=Intermediate, R=Resistant. From Clinical and Laboratory Standards Institute, M100-S5, 015, except ranges in bold/italics established by EUCAST. All ranges have been validated by EUCAST. Target Range susceptibility 1 Comments 1

15 Extended QC EUCAST QC Tables v. 5.0, valid from Quality control strains for detection of resistance mechanisms with disk diffusion on Mueller-Hinton fastidious (MH-F) agar Disk diffusion methodology: Mueller-Hinton agar + 5% defibrinated horse blood and 0 mg/l β-nad, McFarland 0.5, 5% CO, 5±1ºC, 18±h. Read zone edges as the point showing no growth viewed from the front of the plate with the lid removed and with reflected light. Haemophilus influenzae ATCC 4947 (NCTC 1699, CIP , DSM 9999, CCUG 614) Reduced susceptibility to β-lactam agents due to mutations in genes coding for PBP enzymes. (µg) Target Range susceptibility 1 Ampicillin R 6-1 Benzylpenicillin 1 unit R 6-9 Comments s are particularly affected by variation in medium, inoculum and incubation conditions. Inhibition zones with growth of small colonies within the zone are interpreted as 6 mm (no zone). 1 Targets comply with EUCAST clinical breakpoints and are set to ensure that resistance mechanisms are correctly detected. Interpretation according to EUCAST clinical breakpoints: S=Susceptible, I=Intermediate, R=Resistant. Established and validated by repeated testing by EUCAST. 14

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