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Journal of Clinical Microbiology, October 1999, p. 3179-3186, Vol. 37, No. 10
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Multicenter Laboratory Validation of Susceptibility Testing of Mycobacterium tuberculosis against Classical Second-Line and Newer Antimicrobial Drugs by Using the Radiometric BACTEC 460 Technique and the Proportion Method with Solid Media

Gaby E. Pfyffer,1,* Donald A. Bonato,2 Adeleh Ebrahimzadeh,3 Wendy Gross,2 Jacqueline Hotaling,4 John Kornblum,3 Adalbert Laszlo,5 Glenn Roberts,6 Max Salfinger,4 Franziska Wittwer,1 and Salman Siddiqi7

Swiss National Center for Mycobacteria, Department of Medical Microbiology, University of Zurich, Zurich, Switzerland1; Veterans Affairs Medical Center, West Haven, Connecticut2; New York City Department of Health, New York,3 and Wadsworth Center, New York State Department of Health, Albany,4 New York; Laboratory Center for Disease Control, Ottawa, Canada5; Mayo Clinic, Rochester, Minnesota6; and Becton Dickinson Microbiology Systems, Sparks, Maryland7

Received 10 May 1999/Returned for modification 11 July 1999/Accepted 14 July 1999

In a large multicenter study involving six major study sites in the United States, Canada, and Europe, the susceptibilities of 272 Mycobacterium tuberculosis strains to classical second-line antituberculosis (anti-TB) drugs (capreomycin, cycloserine, ethionamide, and kanamycin) and newer compounds (amikacin, clofazimine, ofloxacin, and rifabutin) were determined by the radiometric BACTEC 460 procedure and the conventional proportion method on Middlebrook 7H10 agar. Previously established critical concentrations for classical second-line anti-TB drugs were compared with several concentrations in liquid medium to establish equivalence. MICs of newer compounds determined in liquid medium were either the same or up to four times lower than those determined in agar medium. After establishing critical concentrations (breakpoints) in the extended testing of clinical isolates, we obtained an excellent overall correlation between the two systems, with no errors with amikacin, kanamycin, and ofloxacin and very few major or very major errors with the other drugs; however, for cycloserine, no breakpoint concentration could be recommended due to repeatedly inconsistent results by both methods. Based on these data we conclude that the BACTEC 460 procedure is a simple and rapid method requiring 4 to 8 days on average to generate accurate antimicrobial susceptibility testing (AST) results for eight anti-TB drugs other than those considered primary ones. These data not only fill a major gap of knowledge regarding the critical test concentrations of secondary anti-TB drugs but also provide a baseline for future evaluations of M. tuberculosis AST with the more recently developed, nonradiometric broth-based culture systems.

* Corresponding author. Mailing address: Swiss National Center for Mycobacteria, Department of Medical Microbiology, University of Zurich, Gloriastrasse 30, 8028 Zurich, Switzerland. Phone: 41 1 634 27 86. Fax: 41 1 634 49 18. E-mail: pfyffer{at}immv.unizh.ch.

Journal of Clinical Microbiology, October 1999, p. 3179-3186, Vol. 37, No. 10
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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