The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay is a rapid, cheap, screening test for the in vitro anti-tuberculous activity of chalcones

J Microbiol Methods. 2014 Sep:104:72-8. doi: 10.1016/j.mimet.2014.06.014. Epub 2014 Jun 28.

Abstract

Rapid and reliable drug susceptibility testing facilitates replenishment of the TB drug pipeline in the fight against drug resistant Mycobacterium tuberculosis. This study compared the performance of the MTT and MABA assays on the anti-tuberculous activity of a set of chalcones. Twenty seven chalcones and chromenochalcones were screened against the laboratory strain M. tuberculosis H37Rv, using a microtitre plate MTT assay at 7 days. The MIC for 20 active compounds was subsequently determined using the MABA, MTT and the Macroscopic broth assays at 7, 14 and 21 days. No significant difference in the MICs, or increase in the MICs was observed over time between the MABA (p=0.209) and the MTT (p=0.207) assays, in contrast to the gold standard, the Macroscopic broth assay (p=0.000). The MICs (16 to >128μg/ml) were much higher than the currently used TB drugs. In conclusion, the MTT assay is a cost effective method (R0.06/well) for the rapid in vitro screening of chalcones against M. tuberculosis, producing reliable results in 8 days. The chalcone with a MIC of 16μg/mL shows promise as a potential lead compound and should be investigated further.

Keywords: Alamar blue; Fluorochalcones; M. tuberculosis; MIC; MTT; Macroscopic broth method.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antitubercular Agents / pharmacology*
  • Cell Survival / drug effects
  • Chalcones / pharmacology*
  • Drug Evaluation, Preclinical / instrumentation
  • Drug Evaluation, Preclinical / methods*
  • Formazans / metabolism*
  • Microbial Sensitivity Tests / instrumentation
  • Microbial Sensitivity Tests / methods*
  • Mycobacterium tuberculosis / drug effects*
  • Mycobacterium tuberculosis / growth & development
  • Mycobacterium tuberculosis / metabolism
  • Tetrazolium Salts / metabolism*

Substances

  • Antitubercular Agents
  • Chalcones
  • Formazans
  • Tetrazolium Salts
  • MTT formazan