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

Dumas, Jean-Luc ; van Delden, Christian ; Perron, Karl ; Köhler, Thilo

In: FEMS Microbiology Letters, 2006, vol. 254, no. 2, p. 217-225

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    Title
    • Analysis of antibiotic resistance gene expression in Pseudomonas aeruginosa by quantitative real-time-PCR
    Author
    • Dumas, Jean-Luc. Department of Microbiology and Molecular Medicine, University of Geneva, Geneva, Switzerland
    • van Delden, Christian. Department of Microbiology and Molecular Medicine, University of Geneva, Geneva, Switzerland
    • Perron, Karl. Department of Microbiology and Molecular Medicine, University of Geneva, Geneva, Switzerland
    • Köhler, Thilo. Department of Microbiology and Molecular Medicine, University of Geneva, Geneva, Switzerland
    Document Type
    • Postprint
    OAI-PMH Identifier
    • oai:doc.rero.ch:293861
    Summary
    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 β-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 β-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