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Mechanisms of resistance in nontyphoidal Salmonella enterica strains exhibiting a nonclassical quinolone resistance phenotype.

Antimicrobial agents and chemotherapy (2009-07-15)
Marianne Gunell, Mark A Webber, Pirkko Kotilainen, Andrew J Lilly, Jonathan M Caddick, Jari Jalava, Pentti Huovinen, Anja Siitonen, Antti J Hakanen, Laura J V Piddock
ABSTRACT

Nontyphoidal Salmonella enterica strains with a nonclassical quinolone resistance phenotype were isolated from patients returning from Thailand or Malaysia to Finland. A total of 10 isolates of seven serovars were studied in detail, all of which had reduced susceptibility (MIC > or = 0.125 microg/ml) to ciprofloxacin but were either susceptible or showed only low-level resistance (MIC < or = 32 microg/ml) to nalidixic acid. Phenotypic characterization included susceptibility testing by the agar dilution method and investigation of efflux activity. Genotypic characterization included the screening of mutations in the quinolone resistance-determining regions (QRDR) of gyrA, gyrB, parC, and parE by PCR and denaturing high-pressure liquid chromatography and the amplification of plasmid-mediated quinolone resistance (PMQR) genes qnrA, qnrB, qnrS, qnrD, aac(6')-Ib-cr, and qepA by PCR. PMQR was confirmed by plasmid analysis, Southern hybridization, and plasmid transfer. No mutations in the QRDRs of gyrA, gyrB, parC, or parE were detected with the exception of a Thr57-Ser substitution within ParC seen in all but the S. enterica serovar Typhimurium strains. The qnrA and qnrS genes were the only PMQR determinants detected. Plasmids carrying qnr alleles were transferable in vitro, and the resistance phenotype was reproducible in Escherichia coli DH5alpha transformants. These data demonstrate the emergence of a highly mobile qnr genotype that, in the absence of mutation within topoisomerase genes, confers the nontypical quinolone resistance phenotype in S. enterica isolates. The qnr resistance mechanism enables bacteria to survive elevated quinolone concentrations, and therefore, strains carrying qnr alleles may be able to expand during fluoroquinolone treatment. This is of concern since nonclassical quinolone resistance is plasmid mediated and therefore mobilizable.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Ofloxacin, fluoroquinolone antibiotic
Supelco
Norfloxacin, analytical standard, ≥98% (TLC)
Sigma-Aldrich
Ciprofloxacin, ≥98% (HPLC)
Sigma-Aldrich
Enrofloxacin, ≥99.0%
Sigma-Aldrich
Nalidixic acid, ≥98%
Supelco
Enrofloxacin, VETRANAL®, analytical standard
Supelco
Ciprofloxacin, VETRANAL®, analytical standard
Supelco
Norfloxacin, VETRANAL®, analytical standard
Supelco
Ofloxacin, VETRANAL®, analytical standard
Sigma-Aldrich
Levofloxacin, 98.0-102.0% anhydrous basis (HPLC)