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Mechanisms of Resistance

Topoisomerase II and IV Quinolone Resistance-Determining Regions in Stenotrophomonas maltophilia Clinical Isolates with Different Levels of Quinolone Susceptibility

Sylvia Valdezate, Ana Vindel, Aurora Echeita, Fernando Baquero, Rafael Cantó
Sylvia Valdezate
Servicio de Microbiología, Hospital Ramón y Cajal, Madrid-28034Departamento de Bacteriología, Centro Nacional de Microbiología, Instituto Carlos III, Majadahonda, Madrid-28220, Spain
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Ana Vindel
Departamento de Bacteriología, Centro Nacional de Microbiología, Instituto Carlos III, Majadahonda, Madrid-28220, Spain
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Aurora Echeita
Departamento de Bacteriología, Centro Nacional de Microbiología, Instituto Carlos III, Majadahonda, Madrid-28220, Spain
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Fernando Baquero
Servicio de Microbiología, Hospital Ramón y Cajal, Madrid-28034
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Rafael Cantó
Servicio de Microbiología, Hospital Ramón y Cajal, Madrid-28034
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  • For correspondence: rcanton@hrc.insalud.es
DOI: 10.1128/AAC.46.3.665-671.2002
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ABSTRACT

The quinolone resistance-determining regions (QRDRs) of topoisomerase II and IV genes from Stenotrophomonas maltophilia ATCC 13637 were sequenced and compared with the corresponding regions of 32 unrelated S. maltophilia clinical strains for which ciprofloxacin MICs ranged from 0.1 to 64 μg/ml. GyrA (Leu-55 to Gln-155, Escherichia coli numbering), GyrB (Met-391 to Phe-513), ParC (Ile-34 to Arg-124), and ParE (Leu-396 to Leu-567) fragments from strain ATCC 13637 showed high degrees of identity to the corresponding regions from the phytopathogen Xylella fastidiosa, with the degrees of identity ranging from 85.0 to 93.5%. Lower degrees of identity to the corresponding regions from Pseudomonas aeruginosa (70.9 to 88.6%) and E. coli (73.0 to 88.6%) were observed. Amino acid changes were present in GyrA fragments from 9 of the 32 strains at positions 70, 85, 90, 103, 112, 113, 119, and 124; but there was no consistent relation to higher ciprofloxacin MICs. The absence of changes at positions 83 and 87, commonly involved in quinolone resistance in gram-negative bacteria, was unexpected. The GyrB sequences were identical in all strains, and only one strain (ciprofloxacin MIC, 16 μg/ml) showed a ParC amino acid change (Ser-80→Arg). In contrast, a high frequency (16 of 32 strains) of amino acid replacements was present in ParE. The frequencies of alterations at positions 437, 465, 477, and 485 were higher (P < 0.05) in strains from cystic fibrosis patients, but these changes were not linked with high ciprofloxacin MICs. An efflux phenotype, screened by the detection of decreases of at least twofold doubling dilutions of the ciprofloxacin MIC in the presence of carbonyl cyanide m-chlorophenylhydrazone (0.5 μg/ml) or reserpine (10 μg/ml), was suspected in seven strains. These results suggest that topoisomerases II and IV may not be the primary targets involved in quinolone resistance in S. maltophilia.

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Topoisomerase II and IV Quinolone Resistance-Determining Regions in Stenotrophomonas maltophilia Clinical Isolates with Different Levels of Quinolone Susceptibility
Sylvia Valdezate, Ana Vindel, Aurora Echeita, Fernando Baquero, Rafael Cantó
Antimicrobial Agents and Chemotherapy Mar 2002, 46 (3) 665-671; DOI: 10.1128/AAC.46.3.665-671.2002

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Topoisomerase II and IV Quinolone Resistance-Determining Regions in Stenotrophomonas maltophilia Clinical Isolates with Different Levels of Quinolone Susceptibility
Sylvia Valdezate, Ana Vindel, Aurora Echeita, Fernando Baquero, Rafael Cantó
Antimicrobial Agents and Chemotherapy Mar 2002, 46 (3) 665-671; DOI: 10.1128/AAC.46.3.665-671.2002
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