A rapid detection of multidrug-resistant Mycobacterium tuberculosis by a nitrate reductase assay on blood agar

Çoban, Ahmet Yılmaz; Çaycı, Yeliz Tanrıverdi; Deveci, Aydın; Akgunes, Alper; Uzun, Meltem; Durupinar, Belma

doi:10.1590/s0074-02762011000300022

Cited by 5 publications

(4 citation statements)

References 17 publications

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“…The times required to obtain results were relatively shorter in these studies compared with those in studies performed on LJ medium. Also, susceptibilities of INH and RIF were tested on blood agar media, and it has been suggested that blood agar can be used for the NRA (17). NRA results are negative for M. tuberculosis isolates which do not have nitrate reductase enzymes; however, such isolates are rare.…”

Section: Discussionmentioning

confidence: 99%

Nitrate Reductase Assay for Rapid Detection of Isoniazid, Rifampin, Ethambutol, and Streptomycin Resistance in Mycobacterium tuberculosis: a Systematic Review and Meta-Analysis

et al. 2014

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dColorimetric phenotypic tests recently gained interest because traditional primary drug susceptibility testing of Mycobacterium tuberculosis isolates takes a long time. We used meta-analysis techniques to review the reliability and accuracy of the nitrate reductase assay (NRA), which is one of the most popular colorimetric methods to detect resistance to first-line drugs. Medline, PubMed, ISI Web, Web of Science, and Google Scholar were used to search for studies enrolled in the meta-analysis. The analysis included 35 studies for isoniazid (INH), 38 for rifampin (RIF), and 22 for ethambutol (EMB) and streptomycin (STR). Summary receiver operating characteristic (SROC) curves were applied to summarize diagnostic accuracy. The meta-analyses were performed by the use of Meta-DiSc software (version 1.4) and were focused on sensitivity and specificity values for measurements of accuracy. The pooled sensitivities were 96% for INH, 97% for RIF, 90% for EMB, and 82% for STR. The pooled specificities for INH, RIF, EMB, and STR were 99%, 100%, 98%, and 96%, respectively. The times required to obtain results were between 5 and 28 days by the direct NRA and between 5 and 14 days by the indirect test. In conclusion, the present meta-analysis showed that the NRA is a reliable low-cost rapid colorimetric susceptibility test that can be used for the detection of multidrug-resistant (MDR) tuberculosis, including detection of EMB resistance. However, the test appears to have a relatively low sensitivity for STR and needs further improvement.

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Section: Discussionmentioning

confidence: 99%

Nitrate Reductase Assay for Rapid Detection of Isoniazid, Rifampin, Ethambutol, and Streptomycin Resistance in Mycobacterium tuberculosis: a Systematic Review and Meta-Analysis

et al. 2014

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“…Although this metabolic feature has been used for biochemical identification, recently it has also been employed as a useful growth marker. Other approaches based on nitrate reductase have been studied, such as drug susceptibility test (Kumar et al, 2005;Coban et al, 2011;Dixit et al, 2012) and analysis of the interaction between two or more anti-TB drugs. Despite the wide use of colorimetric methods, the limit of detection and bacterial recovery have not been determined (Il et al, 2013;Rojas-Ponce et al, 2013).…”

Section: Resultsmentioning

confidence: 99%

Fast detection of Mycobacterium tuberculosis in culture-positive sputum samples by nitrate reductase activity

Meneguello

Oliveira

Lima

et al. 2018

Braz. J. Pharm. Sci.

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Microscopy and bacterial culture are the main tools in the diagnosis of tuberculosis. Since the slow growth of Mycobacterium tuberculosis impairs rapid diagnosis strategies, especially in countries where the latter are the only available resources, the ongoing development of new and inexpensive tools based on mycobacterial metabolism optimizing growth detection with preliminary identification is greatly welcome. When compared to the other species from the M. tuberculosis complex, M. tuberculosis is a strong nitrate reducer. Current assay compares the nitrate reductase activity of M. tuberculosis from pulmonary specimens cultivated in nitrate-supplemented media. Fifty-five sputum samples were decontaminated and inoculated in conventional (Middlebrook 7H9, Ogawa Kudoh-OK) and in nitratesupplemented media (Middlebrook 7H9-N, Ogawa Kudoh-N). An aliquot from the media directly reacted with Griess reagent (7H9-N and OK-N) every five days, or transferred to a nitrate substrate solution (7H9, OK). Nitrate to nitrite reduction was considered positive, revealed by the pink color, indicating bacterial growth. As reference method, the Mycobacteria Growth Indicator Tube (MGIT) was used for sensitivity calculations and statistical analysis. 7H9-N and OK-N assays proved to perform better in detecting M. tuberculosis than conventional assays (7H9 and OK). Indeed, broth nitrate-supplemented medium (7H9-N) was comparable to MGIT to detect M. tuberculosis, except in growth detection time. Results show that 7H9-N may be used as an alternative tool particularly in low-income countries since it is a simple and cheap technique, and does not restrict diagnosis to single-source products.

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“…The preparation of Löwenstein-Jensen medium is difficult, and the content is likely to interact with antibiotics [5]. Rapid automated systems (Bactec 460 TB and BACTEC MGIT 960) cannot be obtained by laboratories with limited resources due to equipment requirements and high costs [6].…”

Section: Introductionmentioning

confidence: 99%

Blood Agar Validation for Susceptibility Testing of Isoniazid, Rifampicin, Ethambutol, and Streptomycin to Mycobacterium tuberculosis Isolates

Çoban

2013

PLoS ONE

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In recent studies, it was shown that blood agar can be used at least as effectively as Löwenstein-Jensen medium for growing Mycobacterium tuberculosis. It was also shown that susceptibility testing can be performed on blood agar. Additional validation of blood agar was performed on regional M. tuberculosis isolates from Turkey to determine critical concentrations of isoniazid (INH), rifampicin (RIF), ethambutol (ETM), and streptomycin (STR). In the current study, 40 M. tuberculosis clinical isolates were tested. H37Rv, which is susceptible to all antituberculosis agents, ATCC 35822 (INH-resistant), ATCC 35838 (RIF-resistant), ATCC 35837 (ETM-resistant), and ATCC 35820 (STR-resistant) quality control strains were used as control strains. Proportion method on 7H11 agar was considered as gold standard in the study. MIC values of the control strains and clinical isolates were detected on blood and 7H11 agar. Categorical agreements were 100% for each antibiotic, and essential agreements were 100%, 97.5%, 82.5%, and 95% for INH, RIF, ETM, and STR, respectively. According to the data, 0.2 µg/mL for INH, 1 µg/mL for RIF, 4 µg/mL for ETM, and 2 µg/mL for STR were appropriate breakpoint values for susceptibility testing on blood agar. Blood agar may be recommended for use in both developed and developing countries for the susceptibility testing of M. tuberculosis isolates to primary antituberculosis drugs.

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A rapid detection of multidrug-resistant Mycobacterium tuberculosis by a nitrate reductase assay on blood agar

Cited by 5 publications

References 17 publications

Nitrate Reductase Assay for Rapid Detection of Isoniazid, Rifampin, Ethambutol, and Streptomycin Resistance in Mycobacterium tuberculosis: a Systematic Review and Meta-Analysis

Nitrate Reductase Assay for Rapid Detection of Isoniazid, Rifampin, Ethambutol, and Streptomycin Resistance in Mycobacterium tuberculosis: a Systematic Review and Meta-Analysis

Fast detection of Mycobacterium tuberculosis in culture-positive sputum samples by nitrate reductase activity

Blood Agar Validation for Susceptibility Testing of Isoniazid, Rifampicin, Ethambutol, and Streptomycin to Mycobacterium tuberculosis Isolates

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