空肠弯曲杆菌中喹诺酮类抗生素耐药基因检测与耐药性研究  

Study on the Quinolone Antibiotic Resistance and Its Relative Genes in Campylobacter Jejuni

作　　者：陈云鹏[1] 林雯[1] 邓建平[1] 

机构地区：[1]湖北省黄石市爱康医院,湖北黄石435000

出　　处：《现代检验医学杂志》2015年第2期27-29,共3页Journal of Modern Laboratory Medicine

基　　金：黄石市医疗卫生科技攻关计划项目2012A070-19。

摘　　要：目的：建立一种检测人粪便样本中，空肠弯曲杆菌含喹诺酮类抗生素耐药基因情况的实时荧光 PCR 方法，并对空肠弯曲杆菌对喹诺酮类抗生素耐药性与耐药基因的相关性进行初步研究。方法根据空肠弯曲杆菌对喹诺酮类抗生素耐药基因 gyrA 和 gyrB 序列设计引物，建立对应的实时荧光 PCR 方法，对79例空肠弯曲杆菌进行检测，基因扩增产物经琼脂糖凝胶电泳鉴定。以药敏试验结果作为参照标准，对两种检测方法的结果进行统计学分析，计算实时荧光 PCR 的主要技术指标。结果22例（27.8％）空肠弯曲杆菌的实时荧光 PCR 出现特异性扩增曲线，扩增产物电泳结果显示其中13例携带 gyrA 基因（59.1％），7例携带 gyrB 基因（31.8％），2例同时携带 gyrA 和 gyrB 基因（9.1％）。药敏试验显示26例空肠弯曲杆菌为环丙沙星耐药型（32.9％）。统计学分析显示，两种方法的检测结果差异无统计学意义（χ^2＝1.125，P ＞0.05），一致性较好。实时荧光 PCR 法的灵敏度和特异度分别为76.9％和96.2％，总符合率为89.9％。结论实时荧光PCR 法能够检测空肠弯曲杆菌耐药基因 gyrA 和 gyrB。耐药基因与耐药型之间有所关联，需要进一步研究。Objective To establish a real-time fluorescence PCR method to detect the drug resistance genes of pathogenic Campylobacter jejunum in human stool samples,and investigate the relationship between quinoloneantibiotic resistance and the related genes in Campylobacter jejuni .Methods According to the gyrA and gyrB gene sequences that related with the fluoroquinolone resistance in Campylobacter jejuni ,the primers of the PCR method was designed and synthesized.A rapid real-time fluorescence PCR method to detect the drug resistance genes in Campylobacter jejuni samples was established,and the optimum reaction system and conditions were screened through an optimized approach.The developed method was compared with the classical drug susceptibility assay.Results It was found in the compared results that,there were 8 inconsistent strains of Campylobacter jejuni ,2 of the 8 strains were drug sensitive but contented the drug resistance gene,while 6 strains were drug resistant but had no drug resistant gene.Conclusion The established method can be applied to detect the drug resistance relative genes of gyrA and gyrB in Campylobacter jejuni .There was some correlation between the drug resistance representation and its genotype,but this point requires further studies.

关 键 词：实时荧光PCR 空肠弯曲杆菌 耐药基因 喹诺酮类抗生素 

分 类 号：R378.3[医药卫生—病原生物学] Q786[医药卫生—基础医学]