Rapid test for distinguishing membrane-active antibacterial agents

Journal of Microbiological Methods
Maya Prakash Singh

Abstract

In the search for antibacterial agents with a novel mode-of-action (MOA) many targeted cellular and cell-free assays are developed and used to screen chemical and natural product libraries. Frequently, hits identified by the primary screens include compounds with nonspecific activities that can affect the integrity and function of bacterial membrane. For a rapid dereplication of membrane-active compounds, a simple method was established using a commercially available Live/Dead(R) Bacterial Viability Kit. This method utilized two fluorescent nucleic acid stains, SYTO9 (stains all cells green) and propidium iodide (stains cells with damaged membrane red) for the drug-treated bacterial cells. The cells were then either examined visually by fluorescence microscopy or their fluorescence emissions were recorded using a multi-label plate reader set to measure emissions at two different wavelengths. The ratio of green versus red was compared to a standard curve indicating the percentage of live versus dead bacteria. Nine known antibiotics and 14 lead compounds from various antibacterial screens were tested with results consistent with their MOA.

Citations

Jul 1, 2011·The Journal of Microbiology·Gaëlle LegendreKarine Vallée-Réhel
Apr 6, 2011·Antimicrobial Agents and Chemotherapy·E RubinchikR E W Hancock
Mar 14, 2013·Expert Opinion on Drug Discovery·Herbert A Kirst
Jan 29, 2016·Journal of Medicinal Chemistry·Jordan D GoodreidRobert A Batey
Jun 24, 2015·Annals of the New York Academy of Sciences·Maya A Farha, Eric D Brown
Aug 16, 2014·Journal of Inorganic Biochemistry·Michael L BeetonAlbert Bolhuis
Aug 9, 2017·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Daham JeongSeunho Jung
May 22, 2018·Langmuir : the ACS Journal of Surfaces and Colloids·Farbod AlimohammadiDaniel R Strongin

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