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Antimicrobial Agents and Chemotherapy, November 2003, p. 3430-3434, Vol. 47, No. 11
0066-4804/03/$08.00+0     DOI: 10.1128/AAC.47.11.3430-3434.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Interlaboratory Comparison of Agar Dilution and Etest Methods for Determining the MICs of Antibiotics Used in Management of Neisseria meningitidis Infections

Julio A. Vázquez,1* Luisa Arreaza,1 Colin Block,2 Ingrid Ehrhard,3 Stephen J. Gray,4 Sigrid Heuberger,5 Steen Hoffmann,6 Paula Kriz,7 Pierre Nicolas,8 Per Olcen,9 Anna Skoczynska,10 Lodewijk Spanjaard,11 Paola Stefanelli,12 Muhamed-Kheir Taha,13 and Georgina Tzanakaki14

National Institute of Health Carlos III, Majadahonda (Madrid), Spain,1 Hadassah University Hospital, Jerusalem, Israel,2 National Reference Center for Meningococci, Heidelberg, Germany,3 Meningococcal Reference Unit, Public Health Laboratory, Manchester, United Kingdom,4 Bundesstaatliche Bakterilogisch-Serologischen, Untersuchungsanstalt, Graz, Austria,5 Statens Serum Institute, Copenhagen S, Denmark,6 National Reference Laboratory for Meningococcal Infections, Prague, Czech Republic,7 Unite Meningocoque, WHO Collaborating Center, Marseille, France,8 University Hospital, Örebro, Sweden,9 National Reference Center for Bacterial Meningitis, National Institute of Public Health, Warsaw, Poland,10 Netherlands Reference Laboratory of Bacterial Meningitis, The Netherlands,11 Istituto Superiore di Sanitá, Rome, Italy,12 Institute Pasteur, Paris, France,13 National School of Public Health, Athens, Greece,14

Received 7 February 2003/ Returned for modification 9 June 2003/ Accepted 15 July 2003

Previous studies have shown that there is considerable variation in the methods and media used to determine the susceptibility of Neisseria meningitidis to antimicrobial agents in different countries. In this study, national and regional reference laboratories used a standardized methodology to determine the MICs of antibiotics used in the management of meningococcal infection. Fourteen laboratories participated in the study, determining the susceptibility to penicillin G, rifampin, cefotaxime, ceftriaxone, ciprofloxacin, and ofloxacin of a collection of 17 meningococci, of which 11 strains were previously defined as having intermediate resistance to penicillin (PenI) by sequencing and restriction fragment length polymorphism analysis of the penA gene. The MIC was determined by agar dilution and Etest with Mueller-Hinton agar (MH), MH supplemented with sheep blood (MH+B), and MH supplemented with heated (chocolated) blood. Several laboratories encountered problems obtaining confluent growth with unsupplemented MH. MH+B was considered to give the most congruent and reproducible results among the study laboratories. The modal MIC for MH+B for each antibiotic and method was calculated to define the MIC consensus, allowing assessment of each individual laboratory's data in relation to the others. The agreement in each antibiotic/method/medium combination was defined as the percentage of laboratories with a result within one dilution of the modal result. For the whole study, an agreement of 90.6% was observed between agar dilution and Etest methods. The agreement in each laboratory/antibiotic/method combination ranged from 98.2% to 69.7%, with six laboratories demonstrating agreement higher than 90% and 11 more than 80%. The ability of the laboratories to detect the PenI isolates ranged from 18.2% to 100%. The apparent difficulty in interpreting susceptibility to rifampin, particularly with the Etest method, is very interesting.

* Corresponding author. Mailing address: National Reference Laboratory for Meningococci—Centro Nacional de Microbiología, National Institute of Health Carlos III, 28220 Majadahonda (Madrid), Spain. Phone: 34 915097901. Fax: 34 915097966. E-mail: jvazquez{at}isciii.es.

Antimicrobial Agents and Chemotherapy, November 2003, p. 3430-3434, Vol. 47, No. 11
0066-4804/03/$08.00+0     DOI: 10.1128/AAC.47.11.3430-3434.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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