Detection of ofloxacin resistance by nitrate reductase assay in Mycobacterium tuberculosis isolates from extrapulmonary tuberculosis.

CONTEXT: Increased use of fluoroquinolones to treat community-acquired infections has led to the decreased susceptibility to Mycobacterium tuberculosis. There is a paucity of data on ofloxacin (OFX) resistance detection by nitrate reductase assay (NRA). Hence, the present study was carried out to find the efficacy of NRA for detection of OFX resistance in M. tuberculosis isolated from extrapulmonary tuberculosis (EPTB) cases. AIMS: (1) To compare sensitivity, specificity and median time required to obtain results by NRA with economic variant proportion method (PM) for detection of OFX resistance.(2) To determine the extent of OFX resistance in clinical isolates of M. tuberculosis. SETTINGS AND DESIGN: Seventy-three M. tuberculosis isolates from cases of EPTB were subjected to economic variant of PM for isoniazid, rifampicin and OFX. NRA was done for detection of OFX resistance. SUBJECTS AND METHODS: Seventy-three isolates from clinical samples of suspected EPTB received in the Department of Microbiology were included in the study. Drug susceptibility test was performed on Lowenstein-Jensen medium with and without drugs. STATISTICAL ANALYSIS USED: Of turnaround time was done by Mann-Whitney test on SPSS (version 19, released in 2010, IBM Corp, Armonk NY),P < 0.05. RESULTS: OFX resistance was seen in nine isolates. The sensitivity and specificity of OFX resistance by NRA was 100% and 96.87%, respectively. Median time required to obtain results by NRA was 10 days as compared to 28 days by PM. CONCLUSIONS: NRA is a specific and sensitive method for detection of OFX resistance in resource-restricted settings.

Rapid drug-susceptibility testing of Mycobacterium tuberculosis clinical isolates to first-line antitubercular drugs by nitrate reductase assay: A comparison with proportion method.

OBJECTIVE/BACKGROUND: Early initiation of therapy in patients with tuberculosis is imperative for its control. Conventional methods of susceptibility testing such as the proportion method (PM) require visual detection and counting of colonies that takes up to 6weeks. Rapid and simple phenotypic methods that have been endorsed by the World Health Organization can serve as alternatives. METHODS: In this study, we evaluated the colorimetric nitrate reductase assay, which utilizes the detection of nitrate reduction as an indicator of growth much earlier compared with PM (within 7-14days). The susceptibility of 75 clinical isolates of Mycobacterium tuberculosis to four first-line antitubercular drugs was tested by nitrate reductase assay and compared with the standard PM. In this assay, inoculation was done on both drug-free and drug-containing Löwenstein-Jensen medium containing sodium nitrate. After incubation for 7-14days, reduction to nitrite was taken as an indicator of growth, which was detected by color change on addition of Griess reagent. RESULTS: Agreement between nitrate reductase assay and PM was 100% for rifampicin, 97.30% for isoniazid, 93.30% for streptomycin, and 98.60% for ethambutol. Cost/isolate with this assay was found to be approximately two times lesser than that of PM. All results were obtained in 7-14days by nitrate reductase assay, which was significantly rapid compared with 42days taken for obtaining results by PM. CONCLUSION: Nitrate reductase assay can be used as a rapid and inexpensive method for drug-susceptibility testing of M. tuberculosis for first-line antitubercular drugs without compromising accuracy of standard methods.

[New diagnosis tools for tuberculosis laboratory network in Latin America]. / Consideraciones sobre la implementación de nuevas herramientas diagnósticas en las redes de laboratorios de tuberculosis de Latinoamérica.

Feasibility of rapid and sustainable diagnostic methods that provide useful and timely results to guide the clinical control of tuberculosis patients was analyzed. However, policies guiding the insertion of new diagnostics in the laboratory services that support the tuberculosis control are lacking in developing countries. The introduction of these methods in developing countries laboratories requires rational policies guiding the application of these technologies. In the last few years, some automated systems for culture and molecular testing in laboratory services for tuberculosis diagnosis, which offered accuracy and speed, have been reported. However, their implementation is restricted because of costly resources, logistics and infrastructure. Recently, various economically feasible tests have demonstrated to be applicable in poor-resource labs. The detection of adenosine desaminase (ADA) in pleural fluid is a valuable low-cost approach to the diagnosis of tuberculosis. On the other hand, the microscopic detection of Mycobacterium tuberculosis using thin layer agar is a moderate cost alternative method. Drug susceptibility testing to antituberculous drugs can be expedited by the nitrate reduction assay in tuberculosis laboratories using routine procedures for tuberculosis diagnosis.