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Interlaboratory comparison of agar dilution and Etest methods for determining the MICs of antibiotics used in management of Neisseria meningitidis infections.

Abstract
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 (Pen(I)) 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 Pen(I) isolates ranged from 18.2% to 100%. The apparent difficulty in interpreting susceptibility to rifampin, particularly with the Etest method, is very interesting.
AuthorsJulio A Vázquez, Luisa Arreaza, Colin Block, Ingrid Ehrhard, Stephen J Gray, Sigrid Heuberger, Steen Hoffmann, Paula Kriz, Pierre Nicolas, Per Olcen, Anna Skoczynska, Lodewijk Spanjaard, Paola Stefanelli, Muhamed-Kheir Taha, Georgina Tzanakaki
JournalAntimicrobial agents and chemotherapy (Antimicrob Agents Chemother) Vol. 47 Issue 11 Pg. 3430-4 (Nov 2003) ISSN: 0066-4804 [Print] United States
PMID14576098 (Publication Type: Comparative Study, Journal Article, Research Support, Non-U.S. Gov't)
Chemical References
  • Anti-Bacterial Agents
  • Culture Media
Topics
  • Anti-Bacterial Agents (pharmacology, therapeutic use)
  • Culture Media
  • Europe
  • Meningococcal Infections (drug therapy, microbiology)
  • Microbial Sensitivity Tests (methods)
  • Neisseria meningitidis (drug effects)
  • Polymorphism, Restriction Fragment Length
  • Reference Standards

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