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Journal of Clinical Microbiology, April 1999, p. 1144-1149, Vol. 37, No. 4
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Rapid Film-Based Determination of Antibiotic Susceptibilities of Mycobacterium tuberculosis Strains by Using a Luciferase Reporter Phage and the Bronx Box

Paul F. Riska,1,2 Ya Su,2 Svetoslav Bardarov,3 Lawrence Freundlich,4 Gary Sarkis,5,dagger Graham Hatfull,5 Christian Carrière,3,Dagger Vanaja Kumar,3,§ John Chan,2 and William R. Jacobs Jr.3,*

Department of Medicine, State University of New York Health Science Center at Brooklyn, Brooklyn, New York 112031; Division of Infectious Diseases, Montefiore Medical Center of the Albert Einstein College of Medicine, Bronx, New York 104672; Howard Hughes Medical Institute and Department of Microbiology and Immunology, Albert Einstein College of Medicine,3 and Department of Pathology, Jacobi Medical Center,4 Bronx, New York 10461; and Division of Biological Sciences, University of Pittsburgh, Pittsburgh, Pennsylvania 152605

Received 21 October 1998/Returned for modification 2 December 1998/Accepted 15 January 1999

Detecting antibiotic resistance in Mycobacterium tuberculosis is becoming increasingly important with the global recognition of drug-resistant strains and their adverse impact on clinical outcomes. Current methods of susceptibility testing are either time-consuming or costly; rapid, reliable, simple, and inexpensive methods would be highly desirable, especially in the developing world where most tuberculosis is found. The luciferase reporter phage is a unique reagent well-suited for this purpose: upon infection with viable mycobacteria, it produces quantifiable light which is not observed in mycobacterial cells treated with active antimicrobials. In this report, we describe a modification of our original assay, which allows detection of the emitted light with a Polaroid film box designated the Bronx Box. The technique has been applied to 25 M. tuberculosis reference and clinical strains, and criteria are presented which allow rapid and simple discrimination among strains susceptible or resistant to isoniazid and rifampin, the major antituberculosis agents.

* Corresponding author. Mailing address: Howard Hughes Medical Institute, Albert Einstein College of Medicine of Yeshiva University, 1300 Morris Park Ave., Bronx, NY 10461. Phone: (718) 430-2888. Fax: (718) 518-0366. E-mail: jacobs{at}aecom.yu.edu.

dagger Present address: Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT 06520.

Dagger Present address: Laboratoire de Virologie Moléculaire, Institut de Biologie, Faculté de Médecine de Montpellier, 34060 Montpellier CEDEX, France.

§ Present address: Tuberculosis Research Center, Chetput, Chennai, India 600031.

Journal of Clinical Microbiology, April 1999, p. 1144-1149, Vol. 37, No. 4
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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