title: Preservation of topoisomerase genetic sequences during in vivo and in vitro development of high-level resistance to ciprofloxacin in isogenic Stenotrophomonas maltophilia strains.
abstract: OBJECTIVE
  To ascertain the participation of topoisomerase mutations in the development of ciprofloxacin resistance in isogenic Stenotrophomonas maltophilia mutants.
  
  
  METHODS
  gyrAB and parCE sequences in three paired in vivo isogenic ciprofloxacin-susceptible (MIC range 0.5-4 mg/L) and resistant (16-128 mg/L) S. maltophilia strains (PFGE-characterized) sequentially isolated from three patients, and their corresponding in vitro mutants (ciprofloxacin MIC range 2->128 mg/L), were studied. Efflux phenotype was also investigated.
  
  
  RESULTS
  Despite different quinolone susceptibilities, each paired clinical strain displayed identical gyrAB and parCE sequences as well as their corresponding in vitro mutants. Up to 50% (18/36) of in vitro mutants displayed a positive efflux phenotype when nalidixic acid was combined with MC-207,110, while 6% (2/36) showed the phenotype when exposed to nalidixic acid and reserpine. Carbonyl cyanide m-chlorophenylhydrazone or arsenite failed to alter quinolone MICs.
  
  
  CONCLUSIONS
  The increase of ciprofloxacin MICs in in vivo and in vitro isogenic S. maltophilia mutant strains was not related to quinolone resistance determining region mutations. Highly effective efflux mechanisms might preserve topoisomerase targets from a ciprofloxacin challenge in S. maltophilia.
authors.affiliation: Departamento de Bacteriología, Centro Nacional de Microbiología, Instituto Carlos III, Majadahonda, Madrid, Spain.
authors.forename: Sylvia
authors.initials: S
authors.lastname: Valdezate
authors.forename: Ana
authors.initials: A
authors.lastname: Vindel
authors.forename: Juan Antonio
authors.initials: JA
authors.lastname: Saéz-Nieto
authors.forename: Fernando
authors.initials: F
authors.lastname: Baquero
authors.forename: Rafael
authors.initials: R
authors.lastname: Cantón
mesh_terms: D000328:Adult; D000890:Anti-Infective Agents / Q000494:pharmacology*; D002939:Ciprofloxacin / Q000494:pharmacology*; D027081:DNA Gyrase / Q000235:genetics*; D027101:DNA Topoisomerase IV / Q000235:genetics*; D024881:Drug Resistance, Bacterial; D006801:Humans; D008826:Microbial Sensitivity Tests; D020615:Stenotrophomonas maltophilia / Q000187:drug effects*
chemical_list: D000890:Anti-Infective Agents; D002939:Ciprofloxacin; D027101:DNA Topoisomerase IV; D027081:DNA Gyrase