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Fluoroquinolone resistance in Campylobacter jejuni and Campylobacter coli from poultry and human samples assessed by PCR-restriction fragment length polymorphism assay

机译:PCR-限制性片段长度多态性分析评估家禽和人样品中空肠弯曲杆菌和大肠杆菌弯曲杆菌对氟喹诺酮的耐药性

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摘要

The objective of this study was to determine fluoroquinolone resistance in Campylobacter spp from poultry and human isolates. Forty-one Campylobacter jejuni isolates (30 of poultry origin and 11 of human origin) and 11 Campylobacter coli isolates (10 of human origin and 1 of poultry origin) were examined for ciprofloxacin, norfloxacin, and nalidixic acid resistance using the minimal inhibitory concentration (MIC) method. Thereafter, the isolates were analyzed by PCR–Restriction Fragment Length Polymorphism (RFLP) assay for detection of Thr-86 mutation. Finally, DNA sequencing was performed for confirmation of gyrA gene mutation. A complete correlation was observed between MICs, PCR-RFLP assay, and sequencing. The results revealed high quinolone resistance rates for C. jejuni (100%) and C. coli (100%) isolates obtained from poultry and moderate resistance for C. jejuni (9.1%) and C. coli (40%) samples of human origin. A mutation in codon 86 of the gyrA gene with a Thr-to-Ile substitution is reported to be the main cause of high resistance to quinolones. This mutation can be analyzed by PCR-RFLP assay, which has been proven to be a simple and fast method for the detection of fluoroquinolone resistance in Campylobacter spp.
机译:这项研究的目的是确定家禽和人分离株弯曲杆菌属中氟喹诺酮类药物的耐药性。使用最小抑菌浓度(44%),检查了41株空肠弯曲杆菌分离株(家禽来源30种和人来源11种)和11种大肠杆菌弯曲菌(人来源10种和家禽来源1种)对环丙沙星,诺氟沙星和萘啶酸的耐药性( MIC)方法。此后,通过PCR-限制性片段长度多态性(RFLP)分析法分析分离株,以检测Thr-86突变。最后,进行DNA测序以确认gyrA基因突变。在MIC,PCR-RFLP分析和测序之间观察到完全相关。结果显示,从家禽中获得的空肠弯曲杆菌(100%)和大肠杆菌(100%)分离株对喹诺酮的耐药率高,对人源空肠弯曲杆菌(9.1%)和大肠杆菌(40%)样品的耐药率中等。据报道,具有Thr-Ile取代的gyrA基因密码子86突变是引起喹诺酮类药物高耐药性的主要原因。可以通过PCR-RFLP分析来分析此突变,该方法已被证明是检测弯曲杆菌属中氟喹诺酮耐药性的简便快捷方法。

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