ODELAM, rapid sequence-independent detection of drug resistance in isolates of Mycobacterium tuberculosis

Elife. 2020 May 13;9:e56613. doi: 10.7554/eLife.56613.

Abstract

Antimicrobial-resistant Mycobacterium tuberculosis (Mtb) causes over 200,000 deaths each year. Current assays of antimicrobial resistance need knowledge of mutations that confer drug resistance, or long periods of culture time to test growth under drug pressure. We present ODELAM (One-cell Doubling Evaluation of Living Arrays of Mycobacterium), a time-lapse microscopy-based method that observes individual cells growing into microcolonies. ODELAM enables rapid quantitative measures of growth kinetics in as little as 30 hrs under a wide variety of environmental conditions. We demonstrate ODELAM's utility by identifying ofloxacin resistance in cultured clinical isolates of Mtb and benchmark its performance with standard minimum inhibitory concentration (MIC) assays. ODELAM identified ofloxacin heteroresistance and the presence of drug resistant colony forming units (CFUs) at 1 per 1000 CFUs in as little as 48 hrs. ODELAM is a powerful new tool that can rapidly evaluate Mtb drug resistance in a laboratory setting.

Keywords: Mycobacterium tuberculosis; drug resistance; growth phenotype; heterogeneity; infectious disease; microbiology.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Antibiotics, Antitubercular / pharmacology*
  • Colony Count, Microbial
  • Drug Resistance, Multiple, Bacterial* / genetics
  • Humans
  • Kinetics
  • Microbial Sensitivity Tests*
  • Microscopy, Video*
  • Mycobacterium tuberculosis / drug effects*
  • Mycobacterium tuberculosis / genetics
  • Mycobacterium tuberculosis / growth & development
  • Ofloxacin / pharmacology*
  • Predictive Value of Tests
  • Reproducibility of Results
  • Time-Lapse Imaging*
  • Tuberculosis, Multidrug-Resistant / diagnosis*
  • Tuberculosis, Multidrug-Resistant / drug therapy
  • Tuberculosis, Multidrug-Resistant / microbiology
  • Workflow

Substances

  • Antibiotics, Antitubercular
  • Ofloxacin

Associated data

  • Dryad/10.5061/dryad.b8gtht78q