Demonstration of intrinsic efflux activity of Escherichia coli K-12 AG100 by an automated ethidium bromide method

Miguel Viveiros, Ana Martins, Laura Paixão, Liliana Rodrigues, Marta Martins, Isabel Couto, Eva Fähnrich, Winfried V. Kern, Leonard Amaral

Research output: Contribution to journalArticle

98 Citations (Scopus)

Abstract

Demonstration of efflux of ethidium bromide (EtBr) has been made for over 30 bacterial species, usually by showing enhanced efflux in multidrug-resistant strains that was then abolished by inactivating efflux pumps. Here we present a relatively simple automated method that employs EtBr as an efflux pump substrate for the demonstration of intrinsic efflux activity in Escherichia coli K-12 AG100. The method uses the Rotor-Gene 3000™ instrument for real-time fluorometric measurement of EtBr accumulation under conditions that limit energy (absence of glucose, low temperature) and of EtBr extrusion under optimum conditions. The method can be used for screening compound libraries for efflux inhibiting capacity.

Original languageEnglish
Pages (from-to)458-462
Number of pages5
JournalInternational Journal Of Antimicrobial Agents
VolumeVol. 31
Issue numbern.º 5
DOIs
Publication statusPublished - May 2008

Keywords

  • Automated method
  • Bacterial efflux pumps
  • Efflux pump inhibitors
  • Escherichia coli
  • Ethidium bromide
  • Fluorometry

UN Sustainable Development Goals (SDGs)

  • SDG 3 - Good Health and Well-Being

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