A 96-well plate fluorescence assay for assessment of cellular permeability and active efflux in Salmonella enterica serovar Typhimurium and Escherichia coli

Objectives Multiply antibiotic-resistant (MAR) mutants of Escherichia coli and Salmonella enterica are characterized by reduced susceptibility to several unrelated antibiotics, biocides and other xenobiotics. Porin loss and/or active efflux have been identified as a key mechanisms of MAR. A single r...

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Veröffentlicht in:	Journal of antimicrobial chemotherapy 2010-08, Vol.65 (8), p.1655-1663
Hauptverfasser:	Coldham, Nick G., Webber, Mark, Woodward, Martin J., Piddock, Laura J. V.
Format:	Artikel
Sprache:	eng
Schlagworte:	Anti-Bacterial Agents - metabolism Antibiotics Antibiotics. Antiinfectious agents. Antiparasitic agents Bacterial Proteins - genetics Bacterial Proteins - metabolism Benzimidazoles - metabolism Biological and medical sciences Cell Membrane Permeability Comparative analysis Correlation analysis Drug resistance Drug Resistance, Multiple, Bacterial E coli Escherichia coli Escherichia coli - drug effects Escherichia coli - metabolism Fluorescence Fluorescent Dyes - metabolism Gene Deletion H33342 MAR Medical sciences Membrane Transport Proteins - genetics Membrane Transport Proteins - metabolism Microbial Sensitivity Tests - methods Mutation Pharmacology. Drug treatments Salmonella Salmonella enterica Salmonella Typhimurium Salmonella typhimurium - drug effects Salmonella typhimurium - metabolism Sensitivity and Specificity Staining and Labeling - methods
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Zusammenfassung:	Objectives Multiply antibiotic-resistant (MAR) mutants of Escherichia coli and Salmonella enterica are characterized by reduced susceptibility to several unrelated antibiotics, biocides and other xenobiotics. Porin loss and/or active efflux have been identified as a key mechanisms of MAR. A single rapid test was developed for MAR. Methods The intracellular accumulation of the fluorescent probe Hoechst (H) 33342 (bisbenzimide) by MAR mutants and those with defined disruptions in efflux pump and porin genes was determined in 96-well plate format. Results The accumulation of H33342 was significantly (P
ISSN:	0305-7453 1460-2091
DOI:	10.1093/jac/dkq169