Evaluation of Colorimetric Nitrate Reductase Assay in Liquid Medium for Detection of Resistance to First-line Antitubercular Drugs.


Journal

International journal of mycobacteriology
ISSN: 2212-554X
Titre abrégé: Int J Mycobacteriol
Pays: India
ID NLM: 101615660

Informations de publication

Date de publication:
01 Apr 2024
Historique:
received: 09 03 2024
accepted: 20 05 2024
medline: 25 6 2024
pubmed: 25 6 2024
entrez: 25 6 2024
Statut: ppublish

Résumé

On a global scale, India holds the distinction of having the greatest number of tuberculosis (TB) cases caused by Mycobacterium tuberculosis (MTB) complex. The study aimed at evaluating the sensitivity, specificity, accuracy, cost, rapidity, and feasibility of the performance of the colorimetric nitrate reductase-based antibiotic susceptibility (CONRAS) test against the indirect proportion method (IPM) on Lowenstein-Jensen media as the gold standard. A comparative cross-sectional study was performed on 51 MTB isolates. Fresh subcultures were used for drug susceptibility testing by IPM on the Lowenstein-Jensen medium and the CONRAS method in liquid medium. Quality control for drug susceptibility testing was done using a known sensitive strain of MTB (H37Rv) and strains resistant to both isoniazid (INH) and rifampicin (RIF) - multidrug-resistant (MDR), mono-resistant to RIF, streptomycin (STM), and ethambutol (EMB). Statistical analysis was performed using MedCalc software (Version 20.027). CONRAS, carried out in microfuge tubes, was cost-efficient and easy to perform/interpret with most results being available in 10 days compared to 42 days in the case of IPM. The sensitivity, specificity, and accuracy of RIF and INH were 100%, 97.37%, and 98.04 and 93.33%, 97.59%, and 96.08%, respectively, which translates into an almost perfect agreement between the two methods as indicated by κ value of 0.905 and 0.949, respectively, for the two drugs. The performance of CONRAS was less satisfactory for STM and EMB when compared to IPM. CONRAS may serve as a useful test for the detection of MDR-TB because of its accuracy, low cost, ease of performance/interpretation, and rapidity when compared to IPM on LJ medium. It does not involve the use of expensive reagents and equipment, as is the case with molecular methods like GeneXpert and line probe assay, making it a suitable option for the detection of MDR-TB in resource-poor settings.

Sections du résumé

BACKGROUND BACKGROUND
On a global scale, India holds the distinction of having the greatest number of tuberculosis (TB) cases caused by Mycobacterium tuberculosis (MTB) complex. The study aimed at evaluating the sensitivity, specificity, accuracy, cost, rapidity, and feasibility of the performance of the colorimetric nitrate reductase-based antibiotic susceptibility (CONRAS) test against the indirect proportion method (IPM) on Lowenstein-Jensen media as the gold standard.
METHODS METHODS
A comparative cross-sectional study was performed on 51 MTB isolates. Fresh subcultures were used for drug susceptibility testing by IPM on the Lowenstein-Jensen medium and the CONRAS method in liquid medium. Quality control for drug susceptibility testing was done using a known sensitive strain of MTB (H37Rv) and strains resistant to both isoniazid (INH) and rifampicin (RIF) - multidrug-resistant (MDR), mono-resistant to RIF, streptomycin (STM), and ethambutol (EMB). Statistical analysis was performed using MedCalc software (Version 20.027).
RESULTS RESULTS
CONRAS, carried out in microfuge tubes, was cost-efficient and easy to perform/interpret with most results being available in 10 days compared to 42 days in the case of IPM. The sensitivity, specificity, and accuracy of RIF and INH were 100%, 97.37%, and 98.04 and 93.33%, 97.59%, and 96.08%, respectively, which translates into an almost perfect agreement between the two methods as indicated by κ value of 0.905 and 0.949, respectively, for the two drugs. The performance of CONRAS was less satisfactory for STM and EMB when compared to IPM.
CONCLUSIONS CONCLUSIONS
CONRAS may serve as a useful test for the detection of MDR-TB because of its accuracy, low cost, ease of performance/interpretation, and rapidity when compared to IPM on LJ medium. It does not involve the use of expensive reagents and equipment, as is the case with molecular methods like GeneXpert and line probe assay, making it a suitable option for the detection of MDR-TB in resource-poor settings.

Identifiants

pubmed: 38916391
doi: 10.4103/ijmy.ijmy_69_24
pii: 01929415-202413020-00012
doi:

Substances chimiques

Nitrate Reductase EC 1.7.99.4
Antitubercular Agents 0
Culture Media 0

Types de publication

Journal Article Evaluation Study Comparative Study

Langues

eng

Sous-ensembles de citation

IM

Pagination

191-196

Informations de copyright

Copyright © 2024 Copyright: © 2024 International Journal of Mycobacteriology.

Références

World Health Organization. Global Tuberculosis Report 2023. Geneva: World Health Organization; 2023.
Revendran J, Verma G, Jose JA, Prasana R, Kishan KR, Thanigaiarasu N. Is tuberculosis epidemic riding upon the waves of COVID-19 pandemic? A time-trend study from South India. Int J Mycobacteriol 2023;12:463-6.
Seung KJ, Keshavjee S, Rich ML. Multidrug-resistant tuberculosis and extensively drug-resistant tuberculosis. Cold Spring Harb Perspect Med 2015;5:a017863.
Kammoun S, Smaoui S, Marouane C, Slim L, Messadi-Akrout F. Drug susceptibility testing of Mycobacterium tuberculosis by a nitrate reductase assay applied directly on microscopy-positive sputum samples. Int J Mycobacteriol 2015;4:202-6.
Stephen S, Agrippa K, Majuru Ngoni X, Tariro M, Janet S. Diagnostic performance of STANDARD™ M10 multidrug-resistant tuberculosis assay for detection of Mycobacterium tuberculosis and rifampicin and isoniazid resistance in Zimbabwe. Int J Mycobacteriol 2024;13:22-7.
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.
Palomino JC, Portaels F. Simple procedure for drug susceptibility testing of Mycobacterium tuberculosis using a commercial colorimetic assay. Eur J Clin Microbiol Infect Dis 1999;18:380-3.
Mshana RN, Tadesse G, Abate G, Miörner H. Use of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide for rapid detection of rifampin-resistant Mycobacterium tuberculosis. J Clin Microbiol 1998;36:1214-9.
Coban AY, Akbal AU, Uzun M, Durupinar B. Evaluation of four colourimetric susceptibility tests for the rapid detection of multidrug-resistant Mycobacterium tuberculosis isolates. Mem Inst Oswaldo Cruz 2015;110:649-54.
Moore DA, Evans CA, Gilman RH, Caviedes L, Coronel J, Vivar A, et al. Microscopic-observation drug-susceptibility assay for the diagnosis of TB. N Engl J Med 2006;355:1539-50.
Habous M, E Elimam MA, Kumar R, L Deesi ZA. Evaluation of GeneXpert Mycobacterium tuberculosis/rifampin for the detection of Mycobacterium tuberculosis complex and rifampicin resistance in nonrespiratory clinical specimens. Int J Mycobacteriol 2019;8:132-7.
Dekyong A, Bashir G, Abiroo A, Khan MA, Yasin SB. Multiplex PCR targeting IS6110 and mpb64 genes for detecting extra-pulmonary tuberculosis: A cross-sectional study in a tertiary care center, Kashmir, India. JoMMID 2023;11:78-85.
Gupta A, Sen MR, Mohapatra TM, Anupurba S. Evaluation of the performance of nitrate reductase assay for rapid drug-susceptibility testing of Mycobacterium tuberculosis in North India. J Health Popul Nutr 2011;29:20-5.
Kohli A, Bashir G, Fatima A, Jan A, Wani NU, Ahmad J. Rapid drug-susceptibility testing of Mycobacterium tuberculosis clinical isolates to first-line antitubercular drugs by nitrate reductase assay: A comparison with proportion method. Int J Mycobacteriol 2016;5:469-74.
Coban AY, Birinci A, Ekinci B, Durupinar B. Drug susceptibility testing of Mycobacterium tuberculosis with nitrate reductase assay. Int J Antimicrob Agents 2004;24:304-6.
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.
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.
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.
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.
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.
Affolabi D, Odoun M, Sanoussi N, Martin A, Palomino JC, Kestens L, et al. Rapid and inexpensive detection of multidrug-resistant Mycobacterium tuberculosis with the nitrate reductase assay using liquid medium and direct application to sputum samples. J Clin Microbiol 2008;46:3243-5.
Syre H, Øvreås K, Grewal HM. Determination of the susceptibility of Mycobacterium tuberculosis to pyrazinamide in liquid and solid media assessed by a colorimetric nitrate reductase assay. J Antimicrob Chemother 2010;65:704-12.
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.
Sanchotene KO, von Groll A, Ramos D, Scholante AB, Honscha G, Valença M, et al. Comparative evaluation of the nitrate reductase assay and the resazurin microtitre assay for drug susceptibility testing of Mycobacterium tuberculosis against first line anti-tuberculosis drugs. Braz J Microbiol 2008;39:16-20.
Musa HR, Ambroggi M, Souto A, Angeby KA. Drug susceptibility testing of Mycobacterium tuberculosis by a nitrate reductase assay applied directly on microscopy-positive sputum samples. J Clin Microbiol 2005;43:3159-61.
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 Med Microbiol 2006;55:861-3.
Fonseca Lde S, Vieira GB, Sobral LF, Ribeiro EO, Marsico AG. Comparative evaluation under routine conditions of the nitrate reduction assay, the proportion assay and the MGIT 960 assay for drug susceptibility testing of clinical isolates of Mycobacterium tuberculosis. Mem Inst Oswaldo Cruz 2012;107:142-4.
Ani AE, Daylop YB, Agbaji O, Idoko J. Drug susceptibility test of Mycobacterium tuberculosis by nitrate reductase assay. J Infect Dev Ctries 2009;3:16-9.
Shikama Mde L, Silva RR, Martins MC, Giampaglia CM, Oliveira RS, Silva RF, et al. Rapid detection of resistant tuberculosis by nitrate reductase assay performed in three settings in Brazil. J Antimicrob Chemother 2009;64:794-6.
Gupta M, Shamma Singh NP, Kaur IR. Evaluation of nitrate reductase assay for direct detection of drug resistance in Mycobacterium tuberculosis: Rapid and inexpensive method for low-resource settings. Indian J Med Microbiol 2010;28:363-5.

Auteurs

Munaza Aman (M)

Department of Microbiology, Sher-i-Kashmir Institute of Medical Sciences, Soura, Srinagar, Jammu and Kashmir, India.

Articles similaires

[Redispensing of expensive oral anticancer medicines: a practical application].

Lisanne N van Merendonk, Kübra Akgöl, Bastiaan Nuijen
1.00
Humans Antineoplastic Agents Administration, Oral Drug Costs Counterfeit Drugs

Smoking Cessation and Incident Cardiovascular Disease.

Jun Hwan Cho, Seung Yong Shin, Hoseob Kim et al.
1.00
Humans Male Smoking Cessation Cardiovascular Diseases Female
Humans United States Aged Cross-Sectional Studies Medicare Part C
1.00
Humans Yoga Low Back Pain Female Male

Classifications MeSH