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JAC Advance Access originally published online on June 8, 2007
Journal of Antimicrobial Chemotherapy 2007 60(2):394-397; doi:10.1093/jac/dkm204
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Multiplex PCR for detection of plasmid-mediated quinolone resistance qnr genes in ESBL-producing enterobacterial isolates

Vincent Cattoir1,2, Laurent Poirel1, Vincent Rotimi3, Claude-James Soussy2 and Patrice Nordmann1,*

1 Service de Bactériologie-Virologie, Hôpital de Bicêtre, Assistance Publique/Hôpitaux de Paris, Faculté de Médecine Paris-Sud, Université Paris XI, 78, rue du Général Leclerc, 94275 K.-Bicêtre, France 2 Service de Bactériologie-Virologie-Hygiène, Hôpital Henri Mondor, Assistance Publique/Hôpitaux de Paris, Faculté de Médecine de Créteil, Université Paris XII, Créteil, France 3 Department of Microbiology, Faculty of Medicine, Kuwait University, Kuwait

Received 21 February 2007; returned 11 April 2007; revised 17 April 2007; accepted 11 May 2007

* Corresponding author. Tel: +33-1-45-21-36-32; Fax: +33-1-45-21-63-40; E-mail: nordmann.patrice{at}bct.aphp.fr

Objectives: To develop a rapid and reliable single-tube-based PCR technique for detecting simultaneously the plasmid-mediated quinolone resistance qnrA, qnrB and qnrS genes.

Methods: After multiple alignments, primers were designed to detect known qnr variants (six for qnrA-, six for qnrB- and two for qnrS-like genes). They were used for screening a collection of 64 expanded-spectrum ß-lactamase (ESBL)-producing enterobacterial isolates from Kuwait, collected from 2002 to 2004, as ESBL genes have been often associated with qnr genes. Sequencing was performed to identify qnr and associated ESBL genes.

Results: In optimized conditions, all positive controls (used separately or mixed) confirmed the specificity of the PCR primers. Out of 64 isolates, only 3 isolates were positive for a qnrB-like gene (4.7%), whereas no qnrA-like and qnrS-like gene was detected. A qnrB2 gene was detected in an Enterobacter cloacae K34 (SHV-12+) isolate, whereas qnrB1-like (termed qnrB7) and qnrB6-like (termed qnrB8) genes were identified from E. cloacae K37 (SHV-12+) and Citrobacter freundii K70 (VEB-1b+) isolates, respectively.

Conclusions: We report here a fast and reliable technique for rapid screening of qnr-positive strains to be used for epidemiological surveys. A low prevalence of Qnr determinants among ESBL-producing Enterobacteriaceae was identified in the study with Kuwaiti isolates.

Keywords: qnrA , qnrB , qnrS , Kuwait , Middle East


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