我国部分地区猪养殖场耐药基因mcr-1阳性大肠杆菌的PFGE和RAPD分型对比研究  

PFGE and RAPD Typing of mcr-1-Positive Escherichia coli Isolates on Swine Farms in Different Areas of China

作　　者：仝慧娴 姚晓慧 王舒丰 魏建超[2] 刘珂[2] 邵东华[2] 邱亚峰[2] 马志永[2] 李蓓蓓[2] 刘聚祥[1] TONG Huixian;YAO Xiaohui;WANG Shufeng;WEI Jianchao;LIU Ke;SHAO Donghua;QIU Yafeng;MA Zhiyong;LI Beibei;LIU Juxiang(College of Veterinary Medicine,Hebei Agricultural University,Baoding 071000,China;Shanghai Veterinary Research Institute,CAAS,Shanghai 200241,China)

机构地区：[1]河北农业大学动物医学院,保定071000 [2]中国农业科学院上海兽医研究所,上海200241

出　　处：《中国动物传染病学报》2021年第4期88-94,共7页Chinese Journal of Animal Infectious Diseases

基　　金：上海市青年科技启明星计划(19QA1411200);国家自然基金项目(31672606)。

摘　　要：耐药基因mcr-1严重威胁多黏菌素对多重耐药革兰氏阴性杆菌感染的治疗,该基因起源于动物养殖场,而且具有非常高的流行率。本研究从我国山东、青海、四川、北京、贵州、海南、江西、山西、新疆维吾尔自治区9个省市自治区的33个猪养殖场分离鉴定得到63株mcr-1阳性大肠杆菌,采用脉冲场凝胶电泳分型(PFGE)和随机扩增多态性分析(RAPD)方法来解析上述菌株的亲缘关系,并对两种分子分型方法进行比较。结果显示,我国不同地域猪养殖场中的mcr-1宿主菌主要为大肠杆菌,并且其分子分型呈现多样性特征。63株菌共获得56种PFGE谱型,仅有7组菌株表现出克隆关系;RAPD分型则呈现55种谱型,有7组菌株表现出克隆关系。研究表明两种分型方法均可用于mcr-1阳性大肠杆菌的同源性分析,但PFGE的分辨力、稳定性、重复性都优于RAPD,在对菌株进行亲缘关系分析时推荐使用PFGE。对于缺乏高端设施的实验室,分离株进行分子分型时也可以考虑RAPD。The resistant gene mcr-1 has seriously threatened the treatment of colistin for infections caused by multi-drug resistant Gram-negative bacteria.The gene mcr-1 is present in bacteria of animal origin and shows extremely high prevalence on animal farms.In this study,63 mcr-1-positive Escherichia coli isolates were collected from 33 pig farms in 9 provinces of China,including Shandong,Qinghai,Sichuan,Beijing,Guizhou,Xinjiang,Hainan,Jiangxi and Shanxi.Pulsed fi eld gel electrophoresis(PFGE)and random amplifi ed polymorphic DNA(RAPD)methods were used to analyze the genetic relationships of these mcr-1-positive E.coli isolates.The result showed that the main host bacteria of mcr-1 in Chinese pig farms was E.coli and diverse molecular typing patterns were observed among these isolates.There were 56 PFGE patterns for the 63 mcr-1-positive isolates and only 7 groups showed clonal relationships.For the RAPD typing,55 patterns were observed,among which 7 groups exhibited clonal relationships.Moreover,these two molecular typing methods were compared for their operationality under circumstances.Both typing methods could be used for analysis of genetic relationships of mcr-1-positive E.coli but the resolution stability and repeatability of PFGE were better than those of RAPD.Therefore,PFGE is suggested as the fi rst choice to analyze genetic relationships of bacteria while RAPD can be considered in the laboratories that lack PFGE system.

关 键 词：mcr-1阳性大肠杆菌 同源性分析 脉冲场凝胶电泳分型 随机扩增多态性分析 

分 类 号：S852.612[农业科学—基础兽医学]