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Scientific article
English

Analysis of antibiotic resistance gene expression in Pseudomonas aeruginosa by quantitative real-time-PCR

Published inFEMS microbiology letters, vol. 254, no. 2, p. 217-225
Publication date2006
Abstract

In Pseudomonas aeruginosa many of the clinically relevant resistance mechanisms result from changes in gene expression as exemplified by the Mex drug efflux pumps, the AmpC beta-lactamase and the carbapenem-specific porin OprD. We used quantitative real-time-PCR to analyze the expression of these genes in susceptible and antibiotic-resistant laboratory and clinical strains. In nalB mutants, which overexpress OprM, we observed a four- to eightfold increase in the expression of mexA, mexB, and oprM genes. MexX and mexY genes were induced eight to 12 times in the presence of 2 mg L(-1) tetracycline. The mexC/oprJ and mexE/oprN gene expression levels were increased 30- to 250-fold and 100- to 760-fold in nfxB and nfxC mutants, respectively. We further found that in defined laboratory strains expression levels of ampC and oprD genes paralleled beta-lactamase activity and OprD protein levels, respectively. Our data support the use of quantitative real-time-PCR chain reaction for the analysis of the antimicrobial resistance gene expression in P. aeruginosa.

Keywords
  • Anti-Bacterial Agents/ pharmacology
  • Bacterial Proteins/genetics/ metabolism
  • Drug Resistance, Bacterial/ genetics
  • Gene Expression Regulation, Bacterial
  • Humans
  • Porins/genetics/metabolism
  • Pseudomonas aeruginosa/ drug effects/genetics/metabolism
  • Reverse Transcriptase Polymerase Chain Reaction/ methods
  • Beta-Lactamases/genetics/metabolism
Citation (ISO format)
DUMAS, Jean-Luc Claude et al. Analysis of antibiotic resistance gene expression in Pseudomonas aeruginosa by quantitative real-time-PCR. In: FEMS microbiology letters, 2006, vol. 254, n° 2, p. 217–225. doi: 10.1111/j.1574-6968.2005.00008.x
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ISSN of the journal0378-1097
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