scholarly journals Resazurin Microtiter Assay Plate: Simple and Inexpensive Method for Detection of Drug Resistance in Mycobacterium tuberculosis

2002 ◽  
Vol 46 (8) ◽  
pp. 2720-2722 ◽  
Author(s):  
Juan-Carlos Palomino ◽  
Anandi Martin ◽  
Mirtha Camacho ◽  
Humberto Guerra ◽  
Jean Swings ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Mycobacterium Tuberculosis ◽  
Multidrug Resistant ◽  
Clinical Isolates ◽  
Inexpensive Method ◽  
Antituberculosis Drugs ◽  
Specificity And Sensitivity ◽  
Proportion Method ◽  
Microtiter Assay

ABSTRACT A method for detecting multidrug-resistant Mycobacterium tuberculosis by using a reduction of resazurin is described. Eighty clinical isolates were evaluated against isoniazid and rifampin; results at 7 days were compared with those of the proportion method. Specificity and sensitivity were excellent. The method is simple, inexpensive, and rapid and might be used with other antituberculosis drugs.

Structure–activity relationship of natural and synthetic coumarin derivatives against Mycobacterium tuberculosis

2020 ◽  
Vol 12 (17) ◽  
pp. 1533-1546 ◽  
Author(s):  
Claudia TA Pires ◽  
Regiane BL Scodro ◽  
Diógenes AG Cortez ◽  
Mislaine A Brenzan ◽  
Vera LD Siqueira ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Multidrug Resistant ◽  
Clinical Isolates ◽  
Coumarin Derivatives ◽  
Resistant Tuberculosis ◽  
Diphenyltetrazolium Bromide ◽  
Relationship Of ◽  
Microtiter Assay ◽  
Tuberculosis Activity ◽  
Natural And Synthetic

Aim: Eight coumarin derivatives (1a–h) obtained from natural (-)-mammea A/BB (1) and 13 synthetic coumarins (2–14) had their cytotoxicity and biological activity evaluated against Mycobacterium tuberculosis H37Rv reference strain and multidrug-resistant clinical isolates. Materials & methods: Anti- M. tuberculosis activity was evaluated by resazurin microtiter assay plate, and the cytotoxicity of natural and synthetic products using J774A.1 macrophages by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide. Results: Compounds 1g, 5, 6, 12 and 14 were more active against M. tuberculosis H37Rv and multidrug-resistant clinical isolates with MIC values ranging from 15.6 to 62.5 μg/ml. Conclusion: These results demonstrate that the coumarin derivatives were active against multidrug-resistant clinical isolates, becoming potential candidates to be used in the treatment of resistant tuberculosis.

scholarly journals Molecular Diagnosis of Fluoroquinolone Resistance in Mycobacterium tuberculosis

2014 ◽  
Vol 59 (3) ◽  
pp. 1519-1524 ◽  
Author(s):  
Christine Bernard ◽  
Nicolas Veziris ◽  
Florence Brossier ◽  
Wladimir Sougakoff ◽  
Vincent Jarlier ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
False Negative ◽  
Real Life ◽  
Multidrug Resistant ◽  
Clinical Isolates ◽  
Fluoroquinolone Resistance ◽  
Single Mutation ◽  
Content Type ◽  
Negative Results ◽  
Proportion Method

ABSTRACTAs a consequence of the use of fluoroquinolones (FQ), resistance to FQ has emerged, leading to cases of nearly untreatable and extensively drug-resistant tuberculosis. Mutations in DNA gyrase represent the main mechanism of FQ resistance. A full understanding of the pattern of mutations found in FQ-resistant (FQr) clinical isolates, and of their proportions, is crucial for improving molecular methods for the detection of FQ resistance inMycobacterium tuberculosis. In this study, we reviewed the detection of FQ resistance in isolates addressed to the French National Reference Center for Mycobacteria from 2007 to 2012, with the aim of evaluating the performance of PCR sequencing in a real-life context.gyrAandgyrBsequencing, performed prospectively onM. tuberculosisclinical isolates, was compared for FQ susceptibility to 2 mg/liter ofloxacin by the reference proportion method. A total of 605 isolates, of which 50% were multidrug resistant, were analyzed. The increase in FQrstrains among multidrug-resistant (MDR) strains during the time of the study was alarming (8% to 30%). The majority (78%) of the isolates withgyrAmutations were FQr, whereas only 36% of those withgyrBmutations were FQr. Only 12% of the FQrisolates had a single mutation ingyrB. CombinedgyrAandgyrBsequencing led to >93% sensitivity for detecting resistance. The analysis of the four false-positive and the five false-negative results ofgyrAandgyrBsequencing illustrated the actual limitations of the reference proportion method. Our data emphasize the need for combinedgyrAandgyrBsequencing in the investigation of FQ susceptibility inM. tuberculosisand challenge the validity of the current phenotype-based approach as the diagnostic gold standard for determining FQ resistance.

Activity Of (-)-Camphene Derivatives Against Mycobacterium tuberculosis In Acidic pH

2019 ◽  
Vol 15 ◽  
Author(s):  
Hayalla Corrêa de Carvalho ◽  
Andressa Lorena Ieque ◽  
Tamires Leite Valverde ◽  
Vanessa Pietrowski Baldin ◽  
Jean Eduardo Meneguello ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Culture Medium ◽  
Vero Cells ◽  
Multidrug Resistant ◽  
Tuberculosis H37rv ◽  
Clinical Isolates ◽  
Acidic Ph ◽  
Resistant Tuberculosis ◽  
Low Cytotoxicity ◽  
Microtiter Assay

Background: For more than 60 years, the lack of new anti-tuberculosis drugs and the increase of resistant Mycobacterium tuberculosis lineages exhibit a therapeutic challenge, demanding new options for the treatment of resistant tuberculosis. Objective: Herein, we determined the (i) activities of (-)-camphene and derivatives and (ii) combinatory effect with pyrazinamide (PZA) against Mycobacterium tuberculosis in acidic pH and (iii) cytotoxicity in VERO cells. Methods: The activity of (-)-camphene and 15 derivatives were determined in M. tuberculosis H37Rv in culture medium at pH 6.0 by Resazurin Microtiter Assay Plate (REMA). The combinatory study of three (-)-camphene derivatives with PZA was carried out in seven multidrug-resistant (MDR) clinical isolates by REMA and Checkerboard, respectively. The assay of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium (MTT) bromide in VERO cells was used to determine the derivatives cytotoxicity. Results: Four (-)-Camphene derivatives, (4), (5a) (5d) and (5h), showed reduction in MIC value at pH 6.0 compared to MIC detected at pH 6.8 in M. tuberculosis H37Rv and multidrug resistant clinical isolates. Three (-)-camphene derivatives, (4), (5d) and (5h), showed synergistic effect (FICI ≤ 0.5) combined with PZA and were more selective for M. tuberculosis than VERO cell (selective index from 7.7 to 84.2). Conclusion: Three (-)-camphene derivatives have shown to be promising anti-TB molecule scaffold due to the low MIC values in acidic pH against MDR M. tuberculosis clinical isolates, synergism with PZA and low cytotoxicity.

scholarly journals Second-line anti-tuberculosis drug resistance and its genetic determinants in multidrug-resistant Mycobacterium tuberculosis clinical isolates

2016 ◽  
Vol 49 (3) ◽  
pp. 439-444 ◽  
Author(s):  
Zofia Bakuła ◽  
Agnieszka Napiórkowska ◽  
Michał Kamiński ◽  
Ewa Augustynowicz-Kopeć ◽  
Zofia Zwolska ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Mycobacterium Tuberculosis ◽  
Multidrug Resistant ◽  
Clinical Isolates ◽  
Second Line ◽  
Genetic Determinants

scholarly journals Rapid drug susceptibility testing for Mycobacterium tuberculosis in Thin layer agar media

2013 ◽  
Vol 7 (1) ◽  
pp. 02-06 ◽  
Author(s):  
Habiba Binte Alam ◽  
Md. Ruhul Amin Miah ◽  
S. M. Mostafa Kamal ◽  
Chandan Kumar Roy ◽  
Ahmed Abu Saleh
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Thin Layer ◽  
Rapid Detection ◽  
Drug Susceptibility ◽  
Multidrug Resistant ◽  
Drug Susceptibility Testing ◽  
Clinical Isolates ◽  
Resistant Tuberculosis ◽  
Proportion Method ◽  
Petri Plate

There is a great need to determine the susceptibility of individual Mycobacterium tuberculosis strains as rapidly as possible because emergence of multidrug-resistant and extensively drug-resistant tuberculosis in developing countries. The study was conducted to evaluate the thin layer agar (TLA) media for rapid detection of resistance of M.tuberculosis to rifampicin (RMP) and isoniazid (INH) in clinical isolates and to determine the sensitivity and time to positivity compared to the proportion method. One hundred clinical isolates of M.tuberculosis were studied. For the TLA method, three compartment Petri plate containing 7H11 agar and 7H11 agar with RMP and INH. Results were compared to the proportion method for RMP and INH. The sensitivity for INH and RMP+INH was 85.7 % and 100%. The use of a TLA plate enables the rapid detection of resistance to the two prime anti-tuberculosis drugs RMP and INH in a median time of 9.60 days. TLA was a rapid method for the detection of resistance of M.tuberculosis in the two drugs studied. This faster method is simple to perform, providing an alternative method when more sophisticated techniques are not available in low-resource settings.DOI: http://dx.doi.org/10.3329/bjmm.v7i1.19313 Bangladesh J Med Microbiol 2013; 07(01): 2-6

scholarly journals Resazurin Microtiter Assay Plate Testing of Mycobacterium tuberculosis Susceptibilities to Second-Line Drugs: Rapid, Simple, and Inexpensive Method

2003 ◽  
Vol 47 (11) ◽  
pp. 3616-3619 ◽  
Author(s):  
Anandi Martin ◽  
Mirtha Camacho ◽  
Françoise Portaels ◽  
Juan Carlos Palomino
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Drug Susceptibility ◽  
Multidrug Resistant ◽  
Second Line ◽  
Aminosalicylic Acid ◽  
Resistant Tuberculosis ◽  
Proportion Method ◽  
Susceptibility Tests ◽  
Visual Reading ◽  
Microtiter Assay

ABSTRACT The emergence of multidrug-resistant tuberculosis calls for new, rapid drug susceptibility tests. We have tested 150 Mycobacterium tuberculosis isolates against the second-line drugs ethionamide, kanamycin, capreomycin, ofloxacin, and para-aminosalicylic acid by the colorimetric resazurin microtiter assay and the proportion method. By visual reading, MICs were obtained after 8 days. A very good correlation between results by the colorimetric resazurin microtiter assay and the proportion method was obtained. The colorimetric resazurin microtiter assay is inexpensive, rapid, and simple to perform, and implementation of the assay is feasible for low-resource countries.

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