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JAC Advance Access originally published online on February 24, 2005
Journal of Antimicrobial Chemotherapy 2005 55(4):500-505; doi:10.1093/jac/dki023
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© The Author 2005. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved. For Permissions, please e-mail: journals.permissions{at}oupjournals.org

Comparative evaluation of the nitrate reduction assay, the MTT test, and the resazurin microtitre assay for drug susceptibility testing of clinical isolates of Mycobacterium tuberculosis

Ernesto Montoro1, Dihadenys Lemus1, Miguel Echemendia1, Anandi Martin2, Françoise Portaels2 and Juan Carlos Palomino2,*

1 Laboratorio Nacional de Referencia e Investigaciones en Tuberculosis y Mycobacteria, Instituto de Medicina Tropical ‘Pedro Kouri’, Havana, Cuba; 2 Mycobacteriology Unit, Institute of Tropical Medicine, Nationalestraat 155, 2000 Antwerp, Belgium

* Corresponding author. Tel: +32-3-2476334; Fax: +32-3-2476333; Email: palomino{at}itg.be

Objectives: To evaluate the performance of three rapid low-cost methods for the detection of resistance to first-line drugs in Mycobacterium tuberculosis.

Methods: One hundred M. tuberculosis clinical isolates were tested by the nitrate reductase assay (NRA), the MTT test and the resazurin microtitre assay (REMA), and the results compared with those obtained with the gold standard proportion method (PM) on Löwenstein Jensen medium.

Results: The results using the three methods showed a good sensitivity and specificity between 94% and 100% for the detection of rifampicin and isoniazid resistance. Specificity for ethambutol and streptomycin using MTT and resazurin was low (58–89%). In contrast, NRA showed a good agreement for all first-line drugs tested.

Conclusions: This study shows a high level of agreement of these three low-cost methods compared with the PM for rapid detection of rifampicin and isoniazid resistance. However, more standardization is needed for ethambutol and streptomycin using the MTT test and resazurin microtitre assay. The nitrate reductase assay might represent an inexpensive procedure for rapid detection of resistance to first-line drugs in low-resource countries.

Keywords: antibiotic resistance , colorimetric , rapid methods


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