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

Çoban, Ahmet Yılmaz; Akbal, Ahmet Ugur; Uzun, Meltem; Durupinar, Belma

doi:10.1590/0074-02760150136

Cited by 7 publications

(5 citation statements)

References 15 publications

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“…The MIC values of rifampicin, isoniazid, and natural products interpreted using the REMA method were the same as those value obtained using the CVDA method (Table 1). These results are in accordance with the study conducted by Coban et al. (2015), who reported that REMA has the same values as CVDA, in terms of specificity, sensitivity, positive predictive value, and negative predictive value on susceptibility testing of isoniazid and rifampicin.…”

Section: Discussionsupporting

confidence: 92%

“…Studies on the comparison of REMA and CVDA methods have been carried out previously for detection of M. tuberculosis resistance to primary drugs isoniazid and rifampicin. The results showed that both methods have the same sensitivity and specificity (Coban et al., 2015). However, comparison of both methods for DST anti M. smegmatis has never been conducted, yet.…”

Section: Introductionmentioning

confidence: 99%

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Evaluation of crystal violet decolorization assay and resazurin microplate assay for antimycobacterial screening

Rakhmawatie

Wibawa

Lisdiyanti

et al. 2019

Heliyon

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The main obstacle in antimycobacterial discovery is the extremely slow growth rates of pathogenic mycobacteria that lead to the long incubation times needed in antimycobacterial screening. Some in vitro testings has been developed and are currently available for antimycobacterial screening. The aim of the study was to compare Resazurin Microplate Assay (REMA) and Crystal Violet Decolorization Assay (CVDA) for testing mycobacteria susceptibility to isoniazid and rifampicin as well as for antimycobacterial screening of natural products (NP). Mycobacterium tuberculosis strain H37Rv and Mycobacterium smegmatis strain mc2 155 were used as tested mycobacteria. Serial two-fold dilutions from 0.0625 to 1.0 μg/mL for the isoniazid and rifampicin and from 6.25 to 100.0 μg/mL for the NP A and B were prepared. Tested mycobacteria were then incubated with tested drugs or NPs in each growth medium at 37 °C for 7 days for M. tuberculosis and 3 days for M. smegmatis . MIC values against M. tuberculosis were interpreted 24–48 h after adding resazurin or at least 72 h after adding crystal violet, whereas MIC values against M. smegmatis were interpreted 1 h after adding resazurin or 24 h after adding crystal violet. The MIC values against M. tuberculosis interpreted by REMA were 0.0625, 0.0625, 6.25, and >100 μg/mL for rifampicin, isoniazid, NP A, and NP B, respectively, and those interpreted by CVDA were 0.0625, 0.0625, 6.25, and >100 μg/mL for rifampicin, isoniazid, NP A, and NP B, respectively. Moreover, the MIC values against M. smegmatis interpreted by REMA were 0.0625, >1, 6.25, and 100 μg/mL for rifampicin, isoniazid, NP A, and NP B, respectively, and those interpreted by CVDA were 0.125, >1, 6.25, and >100 μg/mL for rifampicin, isoniazid, NP A, NP B respectively. In conclusion, REMA is faster and easier than CVDA to interpret MIC values, however CVDA produces higher MIC values than REMA for rifampicin and NP B in M. smegmatis susceptibility testing. Therefore, REMA and CVDA can be used for antimycobacterial screening.

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Section: Discussionsupporting

confidence: 92%

Section: Introductionmentioning

confidence: 99%

Evaluation of crystal violet decolorization assay and resazurin microplate assay for antimycobacterial screening

Rakhmawatie

Wibawa

Lisdiyanti

et al. 2019

Heliyon

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“…After this knowledge, CVDA for early detection of MDR-TB was developed by Coban in 2014 . It was reported that agreements for INH and RIF were 94.5-98% and 96.3-98%, respectively ( Coban 2014 , Coban et al 2015) .…”

Section: Discussionmentioning

confidence: 99%

Evaluation of crystal violet decolorization assay for minimal inhibitory concentration detection of primary antituberculosis drugs against Mycobacterium tuberculosis isolates

Çoban

Akbal

Uzun

et al. 2016

Mem. Inst. Oswaldo Cruz

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In this study we evaluated the crystal violet decolorization assay (CVDA) for detection of minimum inhibitory concentration (MIC) of antituberculosis drugs. 53 isolates were tested in this study and 13 of them were multidrug resistant (MDR) isolates. The antibiotics concentrations were 2-0.06 mg/L for isoniazid (INH) and rifampicin (RIF) and were 16-0.25 mg/L for streptomycin (STM) and ethambutol (EMB). Crystal violet (CV-25 mg/L) was added into the microwells on the seventh day of incubation and incubation was continued until decolorization. Decolorization of CV was the predictor of bacterial growth. Overall agreements for four drugs were detected as 98.1%, and the average time was detected as 9.5 ± 0.89 day after inoculation. One isolate for INH and two isolates for STM were determined resistant in the reference method, but susceptible by the CVDA. One isolate was susceptible to EMB by the reference method, but resistant by the CVDA. All results were concordant for RIF. This study shows that CVDA is a rapid, reliable and suitable for determination of MIC values of Mycobacterium tuberculosis. And it can be used easily especially in countries with limited-sources.

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“…The resazurin microtitre assay (REMA) has been developed as a colorimetric and standard method for drug susceptibility testing. The minimum inhibitory concentrations (MICs) were determined according to color changes at the end of incubation [29][30][31]. The strain used, i.e., Mycobacterium tuberculosis H37Rv ATCC 27294 is susceptible to all common antimycobacterial drugs.…”

Section: Antitubercular Activity Assaysmentioning

confidence: 99%

Design, synthesis, antitubercular and antiviral properties of new spirocyclic indole derivatives

et al. 2019

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A series of indole-based spirothiazolidinones have been designed, synthesized and evaluated, in vitro, for their antitubercular, antiviral, antibacterial, and antifungal activities. The structures of the new compounds were established by IR, 1 H NMR, 13 C NMR (proton decoupled, APT, and DEPT), electrospray ionization mass spectrometry, and microanalysis. Compounds bearing a phenyl substituent at position 8 of the spiro ring, exhibited significant antitubercular activity against Mycobacterium tuberculosis H37Rv ATCC 27294 at concentrations of 3.9 and 7.8 µM. Still, some of the tested compounds displayed activity on mycobacteria with MIC values of 16 and 31 µM. Four of the indole-spirothiazolidinone derivatives were found to be moderately active against Punta Toro virus, yellow fever virus or Sindbis virus in Vero cells. The antiviral EC 50 values were in the range of 1.9-12 µM and the selectivity index (ratio of cytotoxic to antivirally effective concentration) was above 10 in some cases. The most potent effect was seen with the compound that is methylated at positions 2 and 8 of the spirothiazolidinone system. Graphic abstract

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

Cited by 7 publications

References 15 publications

Evaluation of crystal violet decolorization assay and resazurin microplate assay for antimycobacterial screening

Evaluation of crystal violet decolorization assay and resazurin microplate assay for antimycobacterial screening

Evaluation of crystal violet decolorization assay for minimal inhibitory concentration detection of primary antituberculosis drugs against Mycobacterium tuberculosis isolates

Design, synthesis, antitubercular and antiviral properties of new spirocyclic indole derivatives

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