Evaluation of nitrate reduction assay, resazurin microtiter assay and microscopic observation drug susceptibility assay for first line antitubercular drug susceptibility testing of clinical isolates of M. tuberculosis

J Microbiol Methods. 2012 Jan;88(1):122-6. doi: 10.1016/j.mimet.2011.11.006. Epub 2011 Nov 26.

Abstract

Background: Drug resistant tuberculosis (TB) is a growing concern worldwide. Early detection of multidrug-resistant Mycobacterium tuberculosis is of primary importance for both patient management and infection control. Optimal method for identifying drug-resistant M. tuberculosis in a timely and affordable way in resource-limited settings is not yet available.

Aim: This study evaluated; nitrate reductase assay (NRA), resazurin microtiter assay (REMA) and microscopic observation drug susceptibility assay (MODS) against the conventional 1% proportion method (PM) for the detection of resistance to first line antitubercular drugs, in M. tuberculosis clinical isolates.

Methods: A total of one hundred and five clinical isolates of M. tuberculosis; 50 pan sensitive and 55 pan resistant were tested with NRA, REMA and MODS. The 1% proportion method on Lowenstein-Jensen medium was used as reference test.

Results: Of all three methods which were tested NRA was found to be most sensitive and specific. Sensitivity for rifampicin resistance detection was 100%, 94.55% and 92.73% by NRA, REMA and MODS respectively. NRA and REMA were found to be 100% specific, while the MODS was 98% specific for detection of rifampicin resistance. Test results with all these methods were obtained within 8-14 days.

Conclusion: Rapid non-conventional and inexpensive methods may serve as a replacement for 1% proportion method in resource limited settings.

Publication types

  • Evaluation Study

MeSH terms

  • Antitubercular Agents / pharmacology*
  • Bacterial Proteins / analysis
  • Bacterial Proteins / metabolism
  • Humans
  • Microbial Sensitivity Tests / methods*
  • Microscopy / methods*
  • Mycobacterium tuberculosis / chemistry
  • Mycobacterium tuberculosis / cytology
  • Mycobacterium tuberculosis / drug effects*
  • Mycobacterium tuberculosis / metabolism
  • Nitrate Reductase / analysis*
  • Nitrate Reductase / metabolism
  • Nitrates / metabolism
  • Oxazines / analysis
  • Sensitivity and Specificity
  • Tuberculosis / microbiology*
  • Xanthenes / analysis

Substances

  • Antitubercular Agents
  • Bacterial Proteins
  • Nitrates
  • Oxazines
  • Xanthenes
  • resazurin
  • Nitrate Reductase