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.
OBJECTIVE
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 Lowenstein 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.
Montoro
Ernesto
E
Laboratorio Nacional de Referencia e Investigaciones en Tuberculosis y Mycobacteria, Instituto de Medicina Tropical 'Pedro Kouri', Havana, Cuba.
Lemus
Dihadenys
D
Echemendia
Miguel
M
Martin
Anandi
A
Portaels
Françoise
F
Palomino
Juan Carlos
JC
D000995:Antitubercular Agents / Q000494:pharmacology*; D024901:Drug Resistance, Multiple, Bacterial; D004977:Ethambutol / Q000494:pharmacology; D005562:Formazans; D007538:Isoniazid / Q000494:pharmacology; D008826:Microbial Sensitivity Tests / Q000379:methods*; D009169:Mycobacterium tuberculosis / Q000187:drug effects* / Q000378:metabolism; D050901:Nitrate Reductase; D009565:Nitrate Reductases / Q000378:metabolism; D010078:Oxazines; D012293:Rifampin / Q000494:pharmacology; D012680:Sensitivity and Specificity; D013307:Streptomycin / Q000494:pharmacology; D013778:Tetrazolium Salts; D014966:Xanthenes
D000995:Antitubercular Agents; D005562:Formazans; D010078:Oxazines; D013778:Tetrazolium Salts; D014966:Xanthenes; C005843:resazurin; C079782:MTT formazan; D004977:Ethambutol; D009565:Nitrate Reductases; D050901:Nitrate Reductase; D007538:Isoniazid; D012293:Rifampin; D013307:Streptomycin