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Journal of Clinical Microbiology, February 2002, p. 553-555, Vol. 40, No. 2
0095-1137/01/$04.00+0     DOI: 10.1128/JCM.40.2.553-555.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Rapid and Inexpensive Drug Susceptibility Testing of Mycobacterium tuberculosis with a Nitrate Reductase Assay

K. A. Kristian Ängeby,1,2* Lisbeth Klintz,1 and Sven E. Hoffner1

Department of Bacteriology, Swedish Institute for Infectious Disease Control, Solna,1 Division of International Health, Karolinska Institute, Stockholm, Sweden2

Received 20 August 2001/ Returned for modification 1 October 2001/ Accepted 15 November 2001

Multidrug-resistant tuberculosis is an increasing public health concern in many parts of the world, especially in low-income countries, where most cases occur. Traditional drug susceptibility testing is either time-consuming, such as the proportion method on solid media, or expensive, such as the BACTEC 460 system. We have evaluated a new nitrate reductase assay (NRA) that depends on the ability of Mycobacterium tuberculosis to reduce nitrate to nitrite. The reduction can be detected using specific reagents, which produce a color change. We tested a panel of 57 M. tuberculosis strains with various resistance patterns. The bacteria were inoculated on Löwenstein-Jensen medium, either without drugs or with rifampin, isoniazid, streptomycin, or ethambutol and with potassium nitrate (KNO3) incorporated. After incubation for 7, 10, or 14 days, the reagents were added and nitrate reduction, indicating growth, could be detected by a color change. Sensitivities to and specificities for drugs as determined by the NRA method compared to those determined by the BACTEC 460 method were 100 and 100% for rifampin, 97 and 96% for isoniazid, 95 and 83% for streptomycin, and 75 and 98% for ethambutol, respectively. The results were in the majority of the cases available in 7 days. The evaluated method is rapid and inexpensive and could correctly identify most resistant and sensitive M. tuberculosis strains. It has the potential to become an interesting alternative to existing methods, such as the proportion and BACTEC methods, particularly in resource-poor settings.

* Corresponding author. Mailing address: Department of Bacteriology, Swedish Institute for Infectious Disease Control, 171 82 Solna, Sweden. Phone: 46-8-457 24 73. Fax 46-8-30 17 97. E-mail: kristian.angeby{at}smi.ki.se.

Journal of Clinical Microbiology, February 2002, p. 553-555, Vol. 40, No. 2
0095-1137/01/$04.00+0     DOI: 10.1128/JCM.40.2.553-555.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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