Utility of nitrate reductase assay for detection of multidrug-resistant Mycobacterium tuberculosis in a low resource setting
Keywords:
Mycobacterium tuberculosis, drug resistance, microbial sensitivity tests, methods, nitrate reductase
Abstract
Introduction. The performance of a drug susceptibility test may change when moving from the research stage to implementation on a population level in actual public health practice.Objective. The performance of a rapid drug susceptibility test was described for detecting multidrug-resistant Mycobacterium tuberculosis when implemented in the routine workflow of a low-resource reference laboratory.
Materials and methods. A prospective study was done comparing the performance of the nitrate reductase assay with the conventional proportion method for rifampicin and isoniazid on 364 isolates were obtained from multidrug-resistant tuberculosis risk patients referred from diffrent Colombian laboratories.
Results. When compared with the proportion method, the nitrate reductase assay sensitivity was 86.8% and 84.9% for rifampicin and isoniazid, respectively, whereas nitrate reductase assay specificity was 100% for isoniazid and rifampicin. Nitrate reductase assay sensitivity was significantly higher when the age of isolate was less than 70 days. A sensitivity of 94.4% dropped to 78.1% for rifampicin resistance for fresh and old isolates, respectively (Fisher exact test, p=0.05). For isoniazid resistance using fresh and old isolates, 94.7% vs.74.3% sensitivities, were achieved (chi square test, p=0.03). The proportion of nitrate reductase assay ambiguous results was significantly higher in multidrug-resistant than in non-multidrug-resistant isolates (17.6% vs. 4.0%, chi square test, p<0.005).
Conclusions. The nitrate reductase assay demonstrated provided reliable results for antibiotic resistance. However, using old cultures leds to a higher proportion of false sensitive results; furthermore, the nitrate reductase assay capability to detect multidrug-resistant tuberculosis decreased due to a higher proportion of non-interpretable results.
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References
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14. Cohn ML, Oda U, Kovitz C, Middlebrook G. Studies on isoniazid and tubercle bacilli. I. The isolation of isoniazid-resistant mutants in vitro. Am Rev Tuberc. 1954;70:465-75.
15. Peizer LR, Widelok D, Klein SC. In vivo observation on Mycobacterium tuberculosis. Their application in the public health laboratory. Am Rev Tuberc. 1956;74:428-37.
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17. Syre H, Phyu S, Sandven P, Bjorvatn B, Grewal HM. Rapid colorimetric method for testing susceptibility of Mycobacterium tuberculosis to isoniazid and rifampin in liquid cultures. J Clin Microbiol. 2003:41:5173-7.
18. Kumar M, Khan IA, Verma V, Kalyan N, Qazi GN. Rapid, inexpensive MIC determination of Mycobacterium tuberculosis isolates by using microplate nitrate reductase assay. Diagn Microbiol Infect Dis. 2005;53:121-4.
19. Kumar M, Khan IA, Verma V, Qazi GN. Microplate nitrate reductase assay versus Alamar Blue assay for MIC determination of Mycobacterium tuberculosis. Int J Tuberc Lung Dis. 2005;9:939-41.
20. Laszlo A, Rahman M, Raviglione M, Bustreo F. Quality assurance programme for drug susceptibility testing of Mycobacterium tuberculosis in the WHO/IUATLD Supranational Laboratory Network: First round of proficiency testing. Int J Tuberc Lung Dis. 1997;1:231-8.
21. Lemus D, Montoro E, Echemendía M, Martin A, Portaels F, Palomino JC. Nitrate reductase assay for detection of drug resistance in Mycobacterium tuberculosis: simple and inexpensive method for low-resource laboratories. J Clin Microbiol. 2006;55:861-3.
22. Shikama M, Silva R, Martins M, Giampaglia C, Oliveira R, Silva R, et al. Rapid detection of resistant tuberculosis by nitrate reductase assay performed in three settings in Brazil. J Antimicrob Chemother. 2009;64:794-6.
23. Rosales S, Pineda-García L, Andino N, Almendarez N, Membreño H, Hoffner SE. Evaluation of the nitrate reductase assay for rapid detection of extensively drug-resistant tuberculosis. Int J Tuberc Lung Dis. 2009;13:1542-9.
24. WHO Strategic and Technical Advisory Group for Tuberculosis (STAG-TB). Report of the Ninth Meeting. 9-11 November 2009. Geneva: WHO; 2009.
2. Palomino JC. Non-conventional and new methods in the diagnosis of tuberculosis. Feasibility and applicability in the field. Eur Respir J. 2005;26:339-50.
3. Palomino JC. Newer diagnostics for tuberculosis and multi-drug resistant tuberculosis. Curr Opin Pulm Med. 2006;12:172-8.
4. Martin A, Panaiotov S, Portaels F, Hoffner S, Palomino JC, Angeby K. The nitrate reductase assay for the rapid detection of isoniazid and rifampicin resistance in Mycobacterium tuberculosis: A systematic review and meta-analysis. J Antimicrob Chemother. 2008;62:56-64.
5. Mengatto L, Chiani Y, Imaz MS. Evaluation of rapid alternative methods for drug susceptibility testing in clinical isolates of Mycobacterium tuberculosis. Mem Inst Oswaldo Cruz. 2006;101:535-42.
6. Small PM, Perkins MD. More rigour need in trials of new diagnostic agents for tuberculosis. Lancet. 2000;356:1048-9.
7. Angeby KA, Klintz L, Hoffner SE. Rapid and inexpensive drug susceptibility testing of Mycobacterium tuberculosis with a nitrate reductase assay. J Clin Microbiol. 2002;40:553-5.
8. Lemus D, Martin A, Montoro E, Portaels F, Palomino J. Rapid alternative methods for detection of rifampicin resistance in Mycobacterium tuberculosis. J Antimicrob Chemother. 2004;54:130-3.
9. Sethi S, Sharma S, Sharma SK, Meharwal SK, Jindal SK, Sharma M. Drug susceptibility of Mycobacterium tuberculosis to primary antitubercular drugs by nitrate reductase assay. Indian J Med Res. 2004;120:468-71.
10. Martin A, Montoro E, Lemus D, Simboli N, Morcillo N, Velasco M, et al. Multicenter evaluation of the nitrate reductase assay for drug resistance detection of Mycobacterium tuberculosis. J Microbiol Methods. 2005;63:145-50.
11. Montoro E, Lemus D, Echemendia M, Martin A, Portaels F, Palomino JC. 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. J Antimicrob Chemother. 2005;55:500-5.
12. Poojary A, Nataraj G, Kanade S, Mehta P, Baveja S. Rapid antibiotic susceptibility testing of Mycobacterium tuberculosis: Its utility in resource poor settings. Indian J Med Microbiol. 2006;24:268-72.
13. NCCLS. Susceptibility testing of Mycobacteria, Nocardia and other aerobic Actinomycetes: Tentative Standard-Second Edition NCCLS document M24-T2 (ISBN 1-56238-423-6), Wayne, Pennsylvania: NCCLS, 2000.
14. Cohn ML, Oda U, Kovitz C, Middlebrook G. Studies on isoniazid and tubercle bacilli. I. The isolation of isoniazid-resistant mutants in vitro. Am Rev Tuberc. 1954;70:465-75.
15. Peizer LR, Widelok D, Klein SC. In vivo observation on Mycobacterium tuberculosis. Their application in the public health laboratory. Am Rev Tuberc. 1956;74:428-37.
16. Organización Panamericana de la Salud. Centro Panamericano de Zoonosis. Bacteriología de la tuberculosis. Sensibilidad del Mycobacterium tuberculosis a las drogas. La identificación de micobacterias. Nota técnica Nº 28. Washington, D.C.: OPS; 1986.
17. Syre H, Phyu S, Sandven P, Bjorvatn B, Grewal HM. Rapid colorimetric method for testing susceptibility of Mycobacterium tuberculosis to isoniazid and rifampin in liquid cultures. J Clin Microbiol. 2003:41:5173-7.
18. Kumar M, Khan IA, Verma V, Kalyan N, Qazi GN. Rapid, inexpensive MIC determination of Mycobacterium tuberculosis isolates by using microplate nitrate reductase assay. Diagn Microbiol Infect Dis. 2005;53:121-4.
19. Kumar M, Khan IA, Verma V, Qazi GN. Microplate nitrate reductase assay versus Alamar Blue assay for MIC determination of Mycobacterium tuberculosis. Int J Tuberc Lung Dis. 2005;9:939-41.
20. Laszlo A, Rahman M, Raviglione M, Bustreo F. Quality assurance programme for drug susceptibility testing of Mycobacterium tuberculosis in the WHO/IUATLD Supranational Laboratory Network: First round of proficiency testing. Int J Tuberc Lung Dis. 1997;1:231-8.
21. Lemus D, Montoro E, Echemendía M, Martin A, Portaels F, Palomino JC. Nitrate reductase assay for detection of drug resistance in Mycobacterium tuberculosis: simple and inexpensive method for low-resource laboratories. J Clin Microbiol. 2006;55:861-3.
22. Shikama M, Silva R, Martins M, Giampaglia C, Oliveira R, Silva R, et al. Rapid detection of resistant tuberculosis by nitrate reductase assay performed in three settings in Brazil. J Antimicrob Chemother. 2009;64:794-6.
23. Rosales S, Pineda-García L, Andino N, Almendarez N, Membreño H, Hoffner SE. Evaluation of the nitrate reductase assay for rapid detection of extensively drug-resistant tuberculosis. Int J Tuberc Lung Dis. 2009;13:1542-9.
24. WHO Strategic and Technical Advisory Group for Tuberculosis (STAG-TB). Report of the Ninth Meeting. 9-11 November 2009. Geneva: WHO; 2009.
How to Cite
1.
López M, Álvarez C. Utility of nitrate reductase assay for detection of multidrug-resistant Mycobacterium tuberculosis in a low resource setting. Biomed. [Internet]. 2011 Mar. 7 [cited 2025 Apr. 4];31(2):258-63. Available from: https://revistabiomedicaorg.biteca.online/index.php/biomedica/article/view/300
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