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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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