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Journal of Clinical Microbiology, March 1999, p. 479-483, Vol. 37, No. 3
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Safe Determination of Susceptibility of Mycobacterium tuberculosis to Antimycobacterial Agents by Flow Cytometry

Andrea V. Moore,1,2 Scott M. Kirk,3 Steven M. Callister,4,5 Gerald H. Mazurek,6 and Ronald F. Schell1,2,7,*

Wisconsin State Laboratory of Hygiene1 and Departments of Medical Microbiology and Immunology2 and Bacteriology,7 University of Wisconsin, Madison, Wisconsin 53706; Microbiology Research Laboratory4 and Section of Infectious Diseases,5 Gundersen Lutheran Medical Center, LaCrosse, Wisconsin 54601; Bio-Rad Laboratories, Hercules, California 945473; and the Centers for Disease Control and Prevention, Atlanta, Georgia 303336

Received 15 October 1998/Returned for modification 18 November 1998/Accepted 4 December 1998

We showed previously that susceptibility testing for Mycobacterium tuberculosis labeled with fluorescein diacetate could be accomplished rapidly by using flow cytometry. However, safety was a major concern because mycobacteria were not killed prior to flow cytometric analysis. In this study, we developed a biologically safe flow cytometric susceptibility test that depends on detection and enumeration of actively growing M. tuberculosis organisms in drug-free and antimycobacterial agent-containing medium. The susceptibilities of 17 clinical isolates of M. tuberculosis to ethambutol, isoniazid, and rifampin were tested by the agar proportion and flow cytometric methods. Subsequently, all flow cytometric susceptibility test samples were inactivated by exposure to paraformaldehyde before analysis with a flow cytometer. Agreement between the results from the two methods was 98%. In addition, the flow cytometric results were available 72 h after the initiation of testing. The flow cytometric susceptibility assay is safe, simple to perform, and more rapid than conventional test methods, such as the BACTEC system and the proportion method.

* Corresponding author. Mailing address: Wisconsin State Laboratory of Hygiene, University of Wisconsin, 465 Henry Mall, Madison, WI 53706. Phone: (608) 262-3634. Fax: (608) 256-3451.

Journal of Clinical Microbiology, March 1999, p. 479-483, Vol. 37, No. 3
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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