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Antimicrob. Agents Chemother. doi:10.1128/AAC.00339-07
Copyright (c) 2007, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

New Plasmid-Mediated Fluoroquinolone Efflux Pump, QepA, Found in an Escherichia coli Clinical Isolate

Department of Bacterial Pathogenesis and Infection Control, National Institute of Infectious Diseases, Tokyo, Japan

^* To whom correspondence should be addressed. Email: yarakawa{at}nih.go.jp.

	Abstract

Plasmid-mediated Qnr and AAC(6')Ib-cr have been recognized as new molecular mechanisms affecting fluoroquinolone (FQ) resistance. An Escherichia coli strain C316 demonstrating resistance to various FQs was isolated in Japan. Resistance level to FQs was augmented in an E. coli CSH2 transconjugant, but PCR failed to detect qnr genes, suggesting the presence of novel plasmid-mediated FQ resistant mechanisms. Susceptibility tests, DNA manipulation, and analyses of gene and its product were performed to characterize the genetic determinant. A novel FQ-resistant gene, qepA, was identified in a plasmid pHPA of E. coli C316, and both qepA and rmtB genes were mediated by a probable transposable element flanked by two copies of IS26. Resistance levels to norfloxacin, ciprofloxacin, and enrofloxacin were significantly elevated in E. coli transformants harboring qepA under AcrAB-TolC deficient condition. The QepA showed a considerable similarity to transporters belonging to the 14-transmembrane segment family of environmental actinomycetes. Effect of carbonyl cyanide m-chlorophenyl hydrazone (CCCP) on accumulation of norfloxacin was assayed in qepA-harboring E. coli transformant. Intracellular accumulation of norfloxacin was decreased in a qepA-expressing E. coli, but this phenomenon was canceled by CCCP. The augmented FQ resistance level acquired by probable intergeneric transfer of a gene encoding for a MFS type efflux pump from some environmental microbes to E. coli was first identified. Surveillance of the qepA-harboring clinical isolates should be encouraged to minimize further dissemination of the kind of plasmid-dependent FQ-resistance determinant among pathogenic microbes.

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