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

Arshad Khan, Dhiman Sarkar

Research output: Contribution to journalArticle

50 Scopus citations

Abstract

This study aimed at developing a whole cell based high throughput screening protocol to identify inhibitors against both active and dormant tubercle bacilli. A respiratory type of nitrate reductase (NarGHJI), which was induced during dormancy, could reflect the viability of dormant bacilli of Mycobacterium bovis BCG in microplate adopted model of in vitro dormancy. Correlation between reduction in viability and nitrate reductase activity was seen clearly when dormant stage inhibitor metronidazole and itaconic anhydride were applied in this in vitro microplate model. Active replicating stage could also be monitored in the same assay by measuring the A620 of the culture. MIC values of 0.08, 0.075, 0.3 and 3.0 μg/ml, determined through monitoring A620 in this assay for rifampin, isoniazid, streptomycin and ethambutol respectively, were well in agreement with previously reported by BACTEC and Bio-Siv assays. S/N ratio and Z' factor for the assay were 8.5 and 0.81 respectively which indicated the robustness of the protocol. Altogether the assay provides an easy, inexpensive, rapid, robust and high content screening tool to search novel antitubercular molecules against both active and dormant bacilli.

Original languageEnglish (US)
Pages (from-to)62-68
Number of pages7
JournalJournal of Microbiological Methods
Volume73
Issue number1
DOIs
StatePublished - Apr 2008

Keywords

  • High throughput screening
  • Tuberculosis
  • Wayne's dormancy model
  • Whole cell based assay

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology
  • Microbiology (medical)

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