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Journal of Clinical Microbiology, May 1998, p. 1214-1219, Vol. 36, No. 5
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Use of 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyl Tetrazolium Bromide for Rapid Detection of Rifampin-Resistant Mycobacterium tuberculosis

Robert N. Mshana,dagger Genet Tadesse, Getahun Abate,Dagger and Håkan Miörner*

Armauer Hansen Research Institute, Addis Ababa, Ethiopia

Received 6 August 1997/Returned for modification 23 September 1997/Accepted 3 February 1998

We describe a test which uses the ability of viable cells to reduce 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) to detect resistance to a bactericidal drug, rifampin, in in vitro-cultured Mycobacterium tuberculosis. The assay shows a linear relationship between the number of viable bacteria and the ability to reduce MTT. Dead mycobacteria were unable to reduce MTT. Rifampin-sensitive M. bovis (BCG) and M. tuberculosis exposed to rifampin showed a rifampin concentration-dependent inhibition of the ability to reduce MTT, while the resistant strains were unaffected. The inhibition of MTT reduction after treatment with rifampin paralleled the reduction in the number of CFU. By using mixing experiments in which the population percentages of rifampin-sensitive and -resistant strains were varied, the assay could detect the presence of rifampin resistance in the mixture when at least 1% of the bacterial population was composed of drug-resistant strains. The assay is cheap, can be visually read, and requires less than 3 days to obtain susceptibility results. The total time required to obtain results, from the time sputum is received in the laboratory, is, in most cases, less than 4 to 5 weeks, which is the time required for primary culture of the bacteria. The MTT assay could, in combination with a test to detect resistance to isoniazid, be a cheap and rapid screening method for multidrug-resistant M. tuberculosis that is affordable even by low-income countries where tuberculosis is a major public health problem.

* Corresponding author. Present address: Department of Mycobacteriology, Statens Serum Institut, Artillerivej 5, DK-2300 Copenhagen S, Denmark. Phone: 45 3268 3720. Fax: 45 3268 3871. E-mail: hmi{at}.ssi.dk.

dagger Present address: Scientific, Technical and Research Commission, Organization of African Unity (OAU), PMB 2359 Lagos, Nigeria.

Dagger Present address: Department of Mycobacteriology, Statens Serum Institut, DK-2300 Copenhagen S, Denmark.

Journal of Clinical Microbiology, May 1998, p. 1214-1219, Vol. 36, No. 5
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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