Comparative evaluation of the microplate nitrate reductase assay and the rezasurin microtitre assay for the rapid detection of multidrug resistant Mycobacterium tuberculosis clinical isolates

Çoban, Ahmet Yılmaz; Uzun, Meltem; Akgunes, Alper; Durupinar, Belma

doi:10.1590/s0074-02762012000500002

Cited by 15 publications

(8 citation statements)

References 7 publications

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“…In this study, the agreement of the MNRA was 96% for INH and 94% for RIF. In the study performed by Coban et al (2012) , the rates for both anti-TB drugs were found to be 100% for the NRA. In another study, Montoro et al (2005) reported that the sensitivity and specificity of the NRA for INH and RIF were 95.6% and 100%, respectively.…”

Section: Discussionmentioning

confidence: 96%

“…Agreements for the REMA were detected to be 98% for both drugs. One study showed that the sensitivities, specificities, PPVs, NPVs and agreements of INH and RIF for the REMA were 100-91.3%, 90.4-100%, 88.5-100%, 100-96.1% and 94.5-97.2%, respectively ( Coban et al 2012 ). Nour et al (2013) also reported that sensitivities, specificities, PPVs, NPVs and agreements of INH and RIF were 100% by this method.…”

Section: Discussionmentioning

confidence: 99%

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Evaluation of four colourimetric susceptibility tests for the rapid detection of multidrug-resistant Mycobacterium tuberculosis isolates

Çoban

Akbal

Uzun

et al. 2015

Mem. Inst. Oswaldo Cruz

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The purpose of this study is to evaluate four rapid colourimetric methods, including the resazurin microtitre assay (REMA), malachite green decolourisation assay (MGDA), microplate nitrate reductase assay (MNRA) and crystal violet decolourisation assay (CVDA), for the rapid detection of multidrug-resistant (MDR) tuberculosis. Fifty Mycobacterium tuberculosis isolates were used in this study. Eighteen isolates were MDR, two isolates were only resistant to isoniazid (INH) and the remaining isolates were susceptible to both INH and rifampicin (RIF). INH and RIF were tested in 0.25 µg/mL and 0.5 µg/mL, respectively. The agar proportion method was used as a reference method. MNRA and REMA were performed with some modifications. MGDA and CVDA were performed as defined in the literature. The agreements of the MNRA for INH and RIF were 96% and 94%, respectively, while the agreement of the other assays for INH and RIF were 98%. In this study, while the specificities of the REMA, MGDA and CVDA were 100%, the specificity of the MNRA was lower than the others (93.3% for INH and 90.9% for RIF). In addition, while the sensitivity of the MNRA was 100%, the sensitivities of the others were lower than that of the MNRA (from 94.1-95%). The results were reported on the seventh-10th day of the incubation. All methods are reliable, easy to perform, inexpensive and easy to evaluate and do not require special equipment.

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Section: Discussionmentioning

confidence: 96%

Section: Discussionmentioning

confidence: 99%

Evaluation of four colourimetric susceptibility tests for the rapid detection of multidrug-resistant Mycobacterium tuberculosis isolates

Çoban

Akbal

Uzun

et al. 2015

Mem. Inst. Oswaldo Cruz

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“…The NRA was usually performed in conventional LJ medium; it had also been performed in liquid media (7H9-S) in 5 studies (12)(13)(14)(15)(16). The times required to obtain results were relatively shorter in these studies compared with those in studies performed on LJ medium.…”

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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“…A resazurin assay was performed to analyze mitochondrial metabolism based on the methods reported by Coban et al [7] and Escribano et al [10]. Resazurin is a non-fluorescent substance that can be reduced by intracellular reducing substances such as nicotinamide adenine dinucleotide phosphate (NADPH), flavin adenine dinucleotide reduced (FADH), flavin mononucleotide reduced (FMNH), and nicotinamide adenine dinucleotide (NADH), as well as by cytochromes [41].…”

Section: Resazurin Assaymentioning

confidence: 99%

Effects of Cold Stimulation on Mitochondrial Activity and VEGF Expression in vitro

et al. 2016

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We aimed to clarify the effects of cold stimulation at various temperatures on mitochondrial activity and vascular endothelial growth factor (VEGF) expression in vitro. Human fibroblast, human mesenchymal stem cell, and rat skeletal muscle myoblast cell lines were used. For each cell type, cells were divided into 4 groups and stimulated in various cold temperatures (0, 4, 17 and 25°C) 3 times for 15 min each by placement on crushed ice or floating on cold water set at each temperature. Control cells were subjected to warm water at 37°C. Factors related to mitochondrial activity, mitochondrial DNA copy numbers, and VEGF expression were analyzed 24 h after the last cold stimulation. In all cell types, significant increases of factors related to mitochondrial activity and mitochondrial DNA copy numbers were seen in the 4°C and 17°C-stimulated cells compared with control cells. In rat skeletal muscle cells stimulated at 4°C, VEGF expression significantly increased compared to the control cells. Our data suggest that cold stimulation at certain temperatures promotes mitochondrial activity, biogenesis and VEGF expression.

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Comparative evaluation of the microplate nitrate reductase assay and the rezasurin microtitre assay for the rapid detection of multidrug resistant Mycobacterium tuberculosis clinical isolates

Cited by 15 publications

References 7 publications

Evaluation of four colourimetric susceptibility tests for the rapid detection of multidrug-resistant Mycobacterium tuberculosis isolates

Evaluation of four colourimetric susceptibility tests for the rapid detection of multidrug-resistant Mycobacterium tuberculosis isolates

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

Effects of Cold Stimulation on Mitochondrial Activity and VEGF Expression in vitro

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