利用添加扩增内标的重组酶聚合酶法检测霍乱弧菌  

Detection of Vibrio Cholerae by Recombinant Enzyme Polymerase with Added Internal Standard

作　　者：庄濠宇 李伟 李窕妍 张峥嵘 温尔英 陈冬娥 周广彪 ZHUANG Haoyu;LI Wei;LI Tiaoyan;ZHANG Zhengrong;WEN Erying;CHEN Dong'e;ZHOU Guangbiao(Jieyang Customs House,Jieyang,Guangdong,522000,China;Meizhou Customs House;Jieyang Market Supervision and Administration Bureau;Shantou Customs District)

机构地区：[1]揭阳海关,广东揭阳522000 [2]梅州海关 [3]揭阳市市场监督管理局 [4]汕头海关

出　　处：《检验检疫学刊》2019年第5期1-5,共5页Journal of Inspection and Quarantine

基　　金：原广东出入境检验检验局科技计划项目(2017GDK48);汕头市科技计划项目(汕府科〔2017〕166号-38)

摘　　要：本文根据霍乱弧菌的ompW基因的保守序列,设计重组酶聚合酶扩增(RPA)特异性引物,构建添加β-action基因为扩增内标的霍乱弧菌RPA-IAC检测方法。从扩增中可得到280 bp的目标基因片段和337 bp的扩增内标片段,该方法特异性良好,检测限与普通PCR一致,灵敏度均可达到0.1 ng/μL。该方法应用测试效果与传统培养方法相当,恒温37℃,40 min即可完成反应,检测效率高,适用于基层及现场检测。According to the conserved sequence of the ompW gene of Vibrio Cholera,a Recombinant enzyme Polymerase Amplification(RPA)specific primer is designed,and a method for detecting the Vibrio cholera RPA-IAC with β-action gene as the internal standard is constructed.A 280 bp target gene fragment and a 337 bp amplified internal standard fragment are obtained from the amplification.The method has good specificity,and the detection limit is consistent with ordinary PCR,and the sensitivity can reach 0.1 ng/μL.The method effect is equivalent to the traditional culture method.The reaction can be completed at a constant temperature of 37℃ for 40 minutes,and the detection has great efficiency.It is a suitable method for the base layer and on-site detection.

关 键 词：霍乱弧菌 重组酶聚合酶扩增 扩增内标 

分 类 号：R446.5[医药卫生—诊断学]