Unit

High-Throughput Assessment of Bacterial Growth Inhibition by Optical Density Measurements

  1. Jennifer Campbell

Published Online: 1 OCT 2010

DOI: 10.1002/9780470559277.ch100115

Current Protocols in Chemical Biology

Current Protocols in Chemical Biology

How to Cite

Campbell, J. 2010. High-Throughput Assessment of Bacterial Growth Inhibition by Optical Density Measurements. Current Protocols in Chemical Biology. 2:195–208.

Author Information

  1. Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts

Publication History

  1. Published Online: 1 OCT 2010
  2. Published Print: OCT 2010

Abstract

The increasing incidence of antibiotic-resistant bacterial infections both in hospitals and in the community intensifies the need for new antibacterial strategies and targets. Although high-throughput screening against live bacteria allows rapid discovery of compounds with growth-inhibitory activities, these efforts have failed to fill the pipeline with the anticipated antibacterial compounds because target identification is often onerous. Recently, a strategy was reported that employs a bacterial growth inhibition assay readout using optical density measurements on paired strains—both a wild-type strain and a pathway-null mutant—to find inhibitors of wild-type bacterial growth that specifically target conditionally essential enzymes in the pathway of interest. Protocols are provided here for determining the robustness of an assay, screening in a high-throughput format, and setting up dose-response curves in paired Staphylococcus aureus strains. However, the protocols can be used to screen for growth-inhibitory compounds in any bacterial strain of interest. Curr. Protoc. Chem. Biol. 2:195-208 © 2010 by John Wiley & Sons, Inc.

Keywords:

  • high-throughput screening;
  • Staphylococcus aureus;
  • optical density;
  • cell-based assay;
  • pathway specific screen;
  • bacterial growth inhibition