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JAC Advance Access originally published online on May 31, 2005
Journal of Antimicrobial Chemotherapy 2005 56(1):220-223; doi:10.1093/jac/dki182
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© The Author 2005. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved. For Permissions, please e-mail: journals.permissions@oupjournals.org

Preservation of topoisomerase genetic sequences during in vivo and in vitro development of high-level resistance to ciprofloxacin in isogenic Stenotrophomonas maltophilia strains

Sylvia Valdezate1, Ana Vindel1, Juan Antonio Saéz-Nieto1, Fernando Baquero2 and Rafael Cantón2,*

1 Departamento de Bacteriología, Centro Nacional de Microbiología, Instituto Carlos III, Majadahonda, Madrid-28220, Spain; 2 Servicio de Microbiología, Hospital Ramón y Cajal, Crta. Colmenar, Km 9, Madrid-28034, Spain

Received 24 January 2005; returned 17 March 2005; revised 28 April 2005; accepted 29 April 2005

* Correspondence address. Servicio de Microbiología, Hospital Universitario Ramón y Cajal, Carretera de Colmenar Km 9.1, 28034-Madrid, Spain; Tel: +34-913368330; Fax: +34-913368809; E-mail: rcanton.hrc{at}salud.madrid.org

Objectives: 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.

Keywords: QRDR , fluoroquinolone resistance , efflux pump inhibitors


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