title: Patterns of quinolone susceptibility in Campylobacter jejuni associated with different gyrA mutations.
abstract: OBJECTIVE
  To investigate the diversity of genetic mutation in the quinolone resistance-determining region (QRDR) of gyrA in clinical isolates and laboratory-derived mutants of Campylobacter jejuni resistant to ciprofloxacin (CipR) and to determine the influence of this mutation on the susceptibility of the organisms to different quinolone antibiotics.
  
  
  METHODS
  Laboratory-derived CipR mutants were obtained from C. jejuni NCTC 11 168 and six quinolone-sensitive faecal isolates (parent prototypes) grown in sub-inhibitory concentrations of ciprofloxacin. Initial mutants found to be CipR were designated 'primary mutants' and subjected to a repeat of this process to select 'secondary mutants' with increased resistance. The susceptibility of the mutants and an additional six clinical isolates of CipR C. jejuni to seven quinolone antibiotics was determined by measuring their MICs. The QRDR of gyrA in all strains was amplified by PCR, sequenced and compared with that of the L04566 C. jejuni gyrA gene.
  
  
  RESULTS
  All six CipR clinical isolates contained a Thr-86-Ile mutation. This was also the commonest mutation found amongst the laboratory derived CipR strains. Other derived mutations in the in vitro derived CipR group included Asp-90-Asn, Thr-86-Ala, and a previously unreported double mutation, Asp-85-Tyr and Thr-86-Ile. Strains with the Thr-86-Ile mutation had the highest MICs to seven different quinolones. CipR strains with other single mutations had a lower range of MICs. There were no additional QRDR mutational changes detected in secondary mutants even where MICs to the fluoroquinolones were higher than in primary mutants.
  
  
  CONCLUSIONS
  Thr-86-Ile mutations were common in both clinical and laboratory derived CipR strains. Other mutations found amongst the latter strains were more sensitive to the fluoroquinolones. Different QRDR changes in gyrA differentially affected the susceptibility of CipR C. jejuni to the various fluoroquinolones.
authors.affiliation: Department of Microbiology (SEALS), Prince of Wales Hospital, Randwick, NSW, Australia. mciverc@sesahs.nsw.gov.au
authors.forename: Christopher
authors.initials: C
authors.lastname: McIver
authors.forename: Tiffany
authors.initials: T
authors.lastname: Hogan
authors.forename: Peter
authors.initials: P
authors.lastname: White
authors.forename: John
authors.initials: J
authors.lastname: Tapsall
mesh_terms: D000890:Anti-Infective Agents / Q000494:pharmacology* / Q000627:therapeutic use; D002169:Campylobacter Infections / Q000188:drug therapy / Q000382:microbiology*; D016123:Campylobacter jejuni / Q000187:drug effects / Q000201:enzymology / Q000235:genetics*; D027081:DNA Gyrase / Q000235:genetics* / Q000378:metabolism; D004252:DNA Mutational Analysis; D004269:DNA, Bacterial / Q000032:analysis; D024881:Drug Resistance, Bacterial / Q000235:genetics*; D006801:Humans; D008826:Microbial Sensitivity Tests; D009154:Mutation; D016133:Polymerase Chain Reaction; D015363:Quinolones / Q000494:pharmacology* / Q000627:therapeutic use
chemical_list: D000890:Anti-Infective Agents; D004269:DNA, Bacterial; D015363:Quinolones; D027081:DNA Gyrase