A simple whole cell based high throughput screening protocol using Mycobacterium bovis BCG for inhibitors against dormant and active tubercle bacilli

Khan, Arshad; Sarkar, Dhiman

doi:10.1016/j.mimet.2008.01.015

Cited by 70 publications

(61 citation statements)

References 29 publications

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“…25 177) using the XTT Reduction Menadione Assay protocol and M. bovis (ATCC No. 35 745) using the Nitrate Reductase assay protocol [19][20][21]. Protocol details are given in Supporting Information.…”

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confidence: 99%

Leucas mollissima, a Source of Bioactive Compounds with Antimalarial and Antimycobacterium Activities

et al. 2015

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!A phytochemical investigation of the acetone extract from the aerial parts of Leucas mollissima afforded one new (−)epi-marmelo lactone, (2 S, 4R, 6 S)-2,6-dimethyl-6 hydroxy-7-ene-4-olide (1), along with five known compounds, schensianol A (2), vanillin (3), β-hydroxy propiovanillone (4), lanost-9(11),25-diene-3β,24β-diol (5), and lanost-9(11),23E(24)-diene-3β,25-diol (6). Similarly, an investigation of the methanol extract of the aerial parts of L. mollissima resulted in the isolation of three known compounds, (+)-syringaresinol (7), anisofolin A (8), and apigenin 7-O-β-D(− 6′′-p-E-coumaroyl)-glucoside (9). Structure elucidation of the isolated compounds was carried out using detailed analysis of 1D and 2D nuclear magnetic resonance. All compounds were evaluated for antimalarial activity against Plasmodium falciparum (3D7) and for antimycobacterium activity against Mycobacterium tuberculosis H37Ra and Mycobacterium bovis. Compound 8 was found to have promising antimalarial activity (IC 50 4.39 ± 0.25 µM), promising antimycobacterium activity [IC 50 4.50 ± 0.75 µM (3.31 µg/mL)] against M. tuberculosis H37Ra and at 100 µg/mL, showed 55.6 % inhibition of M. bovis. Compound 9 showed moderate inhibition of P. falciparum growth (35% inhibition at 10 µM) with respect to the positive control atovaquone and 67.4 % inhibition against M. bovis at 100 µg/mL with respect to the positive control rifampicin.

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Leucas mollissima, a Source of Bioactive Compounds with Antimalarial and Antimycobacterium Activities

et al. 2015

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“…New drug discovery is dependent on whole cell assays to reliably screen for compounds with anti-dormancy, anti-tubercular activities. There is still no specific drug available in the market, which could effectively kill this latent bacillus (Khan & Sarkar, 2008). The obstacle in the development of novel drugs is caused to the lack of a screening system, which can determine inhibitors of latent bacilli of tuberculosis and the limitations of the currently used colonies forming units (CFU) assay (Khan & Sarkar, 2008).…”

Section: Anti-dormant Tubercle Bacilli Assaysmentioning

confidence: 99%

“…There is still no specific drug available in the market, which could effectively kill this latent bacillus (Khan & Sarkar, 2008). The obstacle in the development of novel drugs is caused to the lack of a screening system, which can determine inhibitors of latent bacilli of tuberculosis and the limitations of the currently used colonies forming units (CFU) assay (Khan & Sarkar, 2008). Wayne`s hypoxic model is used for in vitro evaluation of new compounds, but posses low throughput capability (Khan & Sarkar, 2008).…”

Section: Anti-dormant Tubercle Bacilli Assaysmentioning

confidence: 99%

“…The obstacle in the development of novel drugs is caused to the lack of a screening system, which can determine inhibitors of latent bacilli of tuberculosis and the limitations of the currently used colonies forming units (CFU) assay (Khan & Sarkar, 2008). Wayne`s hypoxic model is used for in vitro evaluation of new compounds, but posses low throughput capability (Khan & Sarkar, 2008). Using a M. tuberculosis pFCA-luxAB strain, which is M. tuberculosis H37Rv strain containing a plasmid with an acetamidase promoter driving a bacterial luciferase gene, Cho et al (2007) implemented a high-throughput, luminescence-based low-oxygen-recovery assay for screening of compounds against nonreplicating M. tuberculosis (Cho et al, 2007).…”

Section: Anti-dormant Tubercle Bacilli Assaysmentioning

confidence: 99%

“…Using a M. tuberculosis pFCA-luxAB strain, which is M. tuberculosis H37Rv strain containing a plasmid with an acetamidase promoter driving a bacterial luciferase gene, Cho et al (2007) implemented a high-throughput, luminescence-based low-oxygen-recovery assay for screening of compounds against nonreplicating M. tuberculosis (Cho et al, 2007). An inexpensive alternative is the combination of in vitro model of mycobacterial dormancy with colorimetric methods, recently, Khan & Sarkar (2008) have developed a dormant stage specific antitubercular screening protocol in microplate format using Wayne's hypoxic model and nitrate reductase activity in M. bovis BCG (Bacillus Calmette-Guérin) culture. In addition, Taneja & Tyagi (2007) used resazurin reduction assay to develop screening for searching anti-dormancy and anti-tubercular compounds (Khan & Sarkar, 2008;Taneja & Tyagi, 2007).…”

Section: Anti-dormant Tubercle Bacilli Assaysmentioning

confidence: 99%

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