Scientific article

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

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.

  • 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
Updates (1)
ISSN of the journal0378-1097

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