摘要
针对空肠弯曲菌介导氟喹诺酮类耐药决定区gyrA基因C-257-T位点突变,探讨高分辨率熔解曲线技术对其检测的可行性。选择gyrA基因已经被测序的菌株NCTC11168和ZP11作为高分辨率熔解曲线法的标准阴性和阳性菌株,建立评价高分辨率熔解曲线方法的阴性和阳性标准。利用该技术对已知氟喹诺酮类耐药表型的55株菌株进行检测,结果45株与耐药表型鉴定结果一致,准确率达到81.81%。高分辨率熔解曲线技术检测耐氟喹诺酮类药物空肠弯曲菌gyrA基因C-257-T位点突变具有简单、快速、无污染等优点,有望成为临床检测筛选高耐氟喹诺酮类药物空肠弯曲菌的优选方法。
A high-resolution melting(HRM)assay was developed to detect point mutation C-257-T within QRDR of gyrAgene in Campylobacter jejuni.For development and evaluation of the HRM,C.jejuni NCTC11168 and ZP11were respectively selected as negative and positive standard,and PCR amplicons of gyrA were identified by sequencing.Among 55 additional C.jejuni isolates with known resistance phenotypes,45 of 55fluoroquinolones resistant isolates were identified mutations in gyrAusing HRM and the corresponding specificity was up to 81.81%.HRM is a simple,rapid and pollution-free method for the detection of point mutation C-257-T within QRDR of gyrAgene in C.jejuni,which can be used as an optimized method for screening high-level resistance to fluoroquinolones isolates in clinical samples.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2015年第5期441-447,共7页
Chinese Veterinary Science
基金
国家科技支撑计划项目(2012BAK01B02)
