High-throughput Screening of Chemical Compounds to Elucidate Their Effects on Bacterial Persistence

J Vis Exp. 2021 Feb 23:(168). doi: 10.3791/61597.

Abstract

Bacterial persisters are defined as a small subpopulation of phenotypic variants with the capability of tolerating high concentrations of antibiotics. They are an important health concern as they have been associated with recurrent chronic infections. Although stochastic and deterministic dynamics of stress-related mechanisms are known to play a significant role in persistence, mechanisms underlying the phenotypic switch to/from the persistence state are not completely understood. While persistence factors triggered by environmental signals (e.g., depletion of carbon, nitrogen and oxygen sources) have been extensively studied, the impacts of osmolytes on persistence are yet to be determined. Using microarrays (i.e., 96 well plates containing various chemicals), we have designed an approach to elucidate the effects of various osmolytes on Escherichia coli persistence in a high throughput manner. This approach is transformative as it can be readily adapted for other screening arrays, such as drug panels and gene knockout libraries.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Video-Audio Media

MeSH terms

  • Anti-Bacterial Agents / pharmacology
  • Escherichia coli / drug effects
  • Escherichia coli / genetics
  • Escherichia coli / physiology*
  • High-Throughput Screening Assays / methods*
  • Microarray Analysis
  • Ofloxacin / pharmacology*
  • Reproducibility of Results
  • Solutions

Substances

  • Anti-Bacterial Agents
  • Solutions
  • Ofloxacin