多黏菌素耐药基因mcr-1重组酶介导扩增方法的建立及应用  

Development and Application of Recombinase-aided Amplification Assay(RAA) for Detecting Polymyxin-resistant mcr-1 Gene

作　　者：车勇良[1] 陈秋勇[1] 陈如敬[1] 吴学敏[1] 王隆柏[1] 刘玉涛[1] 周伦江[1] CHE Yong-liang;CHEN Qiu-yong;CHEN Ru-jing;WU Xue-min;WANG Long-bai;LIU Yu-tao;ZHOU Lun-jiang(Institute of Animal Husbandry and Veterinary Medicine,Fujian Academy of Agricultural Sciences/Fujian Animal Diseases Control Techology Development Center,Fuzhou Fujian,350013,China)

机构地区：[1]福建省农业科学院畜牧兽医研究所/福建省畜禽疫病防治工程技术研究中心,福建福州350013

出　　处：《动物医学进展》2022年第8期46-49,共4页Progress In Veterinary Medicine

基　　金：国家自然科学基金项目(31872502);福建省公益类科研院所专项(2019R1026-13);福建省农业科学院科研项目(AGY2018-6)。

摘　　要：根据多黏菌素耐药基因mcr-1的保守序列,设计合成一对重组酶介导扩增方法(RAA)特异性引物和一条FAM标记的荧光探针,优化扩增条件,建立了检测多黏菌素耐药基因mcr-1的RAA方法,验证其特异性和敏感性,并应用于临床检测。结果显示,建立的RAA方法只需21 min就能出结果,能特异性地检测多黏菌素耐药基因mcr-1,最低能检测出10~2的mcr-1阳性质粒拷贝数,也可很好地检测出临床样品中的mcr-1基因。4株临床分离大肠埃希氏菌株样品,有2株mcr-1基因阳性,2株为mcr-1基因阴性。In order to well and quickly detect mcr-1 gene resistant-polymyxin, the recombinase-aided amplification assay(RAA) was established.According to the conserved sequence of mcr-1 gene, a pair of specific primiers and a FAM labeled probe were designed and synthesized.By optimized the amplification condition, a RAA method for detecting mcr-1 gene was preliminary established.The specificity and sensitivity of the RAA were verified, and the RAA method was applied to detect clinical samples.The results showed that detecting time was in 21 min, and the RAA method could only detect the mcr-1 gene, and the minimum detecting number of mcr-1 positive plasmid copies was 10~2.2 strain bacteria in the 4 strian bacteria from clinical sample were mcr-1 gene positive, 2 strain bacteria were mcr-1 gene negative by RAA.

关 键 词：多黏菌素 耐药基因 mcr-1 重组酶介导扩增方法 

分 类 号：S859.7[农业科学—临床兽医学]