title: High-throughput assessment of bacterial growth inhibition by optical density measurements.
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.
authors.affiliation: Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Campbell
keywords: Staphylococcus aureus; bacterial growth inhibition; cell‐based assay; high‐throughput screening; optical density; pathway specific screen