Streptococcus parauberis 의 퀴놀론 내성 증가와 Topoisomerase 유전자에서의 돌연변이 신속 분석
Increased Resistance to Quinolones in Streptococcus parauberis and Development of a Rapid Assay for Detecting Mutations in Topoisomerase Genes
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To investigate the acquisition of quinolone resistance, we examined mutations in the quinolone resistance-determining region (QRDR) of type II topoisomerase genes in ciprofloxacin (CIP)-resistant clinical isolates and in vitro mutants of Streptococcus parauberis . The CIP-resistant clinical isolates had one base change responsible for a Ser-79→Thr in the QRDR of par C. However, the CIP-resistant in vitro mutants had an altered QRDR of par C (Ser-79→Ile) that differed from that of the isolates. None of the CIP-resistant S. parauberis clinical isolates or in vitro mutants exhibited amino acid changes in gyr A or gyr B. However, even though involvement in the increased resistance was not clear, an Arg-449→Ser mutation outside of the QRDR of par E was detected in CIP-resistant mutant 2P1. These results suggest that the topoisomerase IV gene, par C (and possibly par E, as well), is the primary ciprofloxacin target in S. parauberis . Additionally we established a high-resolution melting (HRM) assay capable of detecting the dominant mutation in four type II topoisomerase genes conferring ciprofloxacin resistance. These rapid and reliable assays may provide a convenient method of surveillance for genetic mutations conferring antibiotic resistance.
Abstract
To investigate the acquisition of quinolone resistance, we examined mutations in the quinolone resistance-determining region (QRDR) of type II topoisomerase genes in ciprofloxacin (CIP)-resistant clinical isolates and in vitro mutants of Streptococcus parauberis . The CIP-resistant clinical isolates had one base change responsible for a Ser-79→Thr in the QRDR of par C. However, the CIP-resistant in vitro mutants had an altered QRDR of par C (Ser-79→Ile) that differed from that of the isolates. None of the CIP-resistant S. parauberis clinical isolates or in vitro mutants exhibited amino acid changes in gyr A or gyr B. However, even though involvement in the increased resistance was not clear, an Arg-449→Ser mutation outside of the QRDR of par E was detected in CIP-resistant mutant 2P1. These results suggest that the topoisomerase IV gene, par C (and possibly par E, as well), is the primary ciprofloxacin target in S. parauberis . Additionally we established a high-resolution melting (HRM) assay capable of detecting the dominant mutation in four type II topoisomerase genes conferring ciprofloxacin resistance. These rapid and reliable assays may provide a convenient method of surveillance for genetic mutations conferring antibiotic resistance.
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