title: Development and evaluation of a multiplex PCR for eight plasmid-mediated quinolone-resistance determinants.
abstract: The objective of this study was to develop and validate an expanded multiplex PCR assay for the simultaneous detection of eight plasmid-mediated quinolone-resistance determinants in Enterobacteriaceae. Primers were designed to amplify conserved fragments of qnrABCDS, qepA, oqxAB and aac(6')-Ib-cr genes and were optimized in uniplex and multiplex PCR assays with control template DNA. The assay was used to determine the prevalence of plasmid-mediated quinolone resistance (PMQR) genes in 174 ciprofloxacin-resistant and 43 ciprofloxacin-susceptible extraintestinal pathogenic Escherichia coli isolates. Each resistance gene could be detected alone and in combination. PMQR determinants were detected in 65 ciprofloxacin-resistant isolates (37 %) and one ciprofloxacin-susceptible isolate (2 %). Prevalences of the identified determinants were: aac(6')-Ib-cr, 34.5 %; qnrS, 1.1 %; qepA, 1.1 %; and oqxAB, 0.6 %. In conclusion, we developed an eight-target multiplex PCR for the accurate detection of PMQR genes and confirmed that PMQR prevalence remains low among human Escherichia coli clinical isolates in the UK.
authors.affiliation: Antimicrobial Resistance and Healthcare Associated Infections Reference Laboratory, Public Health England, London NW9 5EQ, UK.|Antimicrobial Research Group, Centre for Immunology and Infectious Disease, Blizard Institute, Barts and the London School of Medicine and Dentistry, Queen Mary, University of London, London E1 2AT, UK.
authors.forename: H
authors.initials: H
authors.lastname: Ciesielczuk
authors.affiliation: Department of Life Sciences, Whitelands College, University of Roehampton, Holybourne Avenue, London SW15 4JD, UK.|Antimicrobial Research Group, Centre for Immunology and Infectious Disease, Blizard Institute, Barts and the London School of Medicine and Dentistry, Queen Mary, University of London, London E1 2AT, UK.
authors.forename: M
authors.initials: M
authors.lastname: Hornsey
authors.affiliation: Singapore General Hospital, Singapore 169037, Singapore.|Antimicrobial Research Group, Centre for Immunology and Infectious Disease, Blizard Institute, Barts and the London School of Medicine and Dentistry, Queen Mary, University of London, London E1 2AT, UK.
authors.forename: V
authors.initials: V
authors.lastname: Choi
authors.affiliation: Antimicrobial Resistance and Healthcare Associated Infections Reference Laboratory, Public Health England, London NW9 5EQ, UK.|Antimicrobial Research Group, Centre for Immunology and Infectious Disease, Blizard Institute, Barts and the London School of Medicine and Dentistry, Queen Mary, University of London, London E1 2AT, UK.
authors.forename: N
authors.initials: N
authors.lastname: Woodford
authors.affiliation: Antimicrobial Research Group, Centre for Immunology and Infectious Disease, Blizard Institute, Barts and the London School of Medicine and Dentistry, Queen Mary, University of London, London E1 2AT, UK.
authors.forename: D W
authors.initials: DW
authors.lastname: Wareham
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D002939:Ciprofloxacin / Q000494:pharmacology*; D004352:Drug Resistance, Microbial / Q000235:genetics*; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics*; D004927:Escherichia coli Infections / Q000175:diagnosis / Q000188:drug therapy / Q000235:genetics; D029968:Escherichia coli Proteins / Q000235:genetics; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D010957:Plasmids / Q000235:genetics*; D015995:Prevalence
chemical_list: D000900:Anti-Bacterial Agents; D029968:Escherichia coli Proteins; D002939:Ciprofloxacin