蓝舌病、口蹄疫、小反刍兽疫和水泡性口炎多重PCR检测方法的建立  

Development of Multiplex PCR Method for Detection of BTV,FMDV,PPRV and VSV

作　　者：秦敏[1] 邹丰才[1] 杨云庆[2] 祝贺[2] 聂福平[3] 王煜[3] 杨俊[3] 叶玲玲[2] 周晓黎[2] 艾军[2] 

机构地区：[1]云南农业大学,云南昆明650201 [2]云南出入境检验检疫局,云南昆明650228 [3]重庆出入境检验检疫局,四川重庆400020

出　　处：《动物医学进展》2015年第9期18-22,共5页Progress In Veterinary Medicine

基　　金：国家质检总局公益性项目(201310093);国家科技支撑计划项目(2013BAD12B03)

摘　　要：为建立一种检测并鉴别蓝舌病病毒、口蹄疫病毒、小反刍兽疫病毒和水泡性口炎病毒感染的方法,针对蓝舌病病毒NS3基因、口蹄疫病毒3D基因、小反刍兽疫病毒N基因和水泡性口炎病毒N基因序列设计引物,优化反应体系和扩增条件,建立了一种同时检测4种病毒的多重PCR方法。对建立的多重PCR检测方法的特异性及敏感性进行检验,结果表明建立的多重PCR检测方法敏感性强、特异性良好,对4种病毒的最低检出限分别为PPRV 103 copies/μL、BTV 103 copies/μL、VSV 103 copies/μL和FMDV 102 copies/μL。本方法的建立对临床感染蓝舌病等4种病毒的病畜进行快速检测具有十分重要的意义。In order to establish a method for detection and identification of BTV,FMDV,PPRV and VSV infection,four pairs of primers based on the gene sequences of BTV NS3, FMDV 3D,PPRV N and VSV N were designed to find out a multiplex PCR method which can test the four viruses at the same time. The amplification system and conditions were optimized. According to the specificity and sensitivity tests of the multiplex PCR method,the results showed that the newly-established method has high sensitivity and spe- cificity,the lowest levels of the four viruses are copies/μL,FMDV 102 copies/μL respectively. This PPRV 103 copies/μL, BTV 103 copies/μL, VSV 103 method stuy not only can identify the four kinds of vi-ruses, such as BTV,GPV, SPV, LSDV, and can also detect the animal infected with four viruses.

关 键 词：蓝舌病 口蹄疫 小反刍兽疫 水泡性口炎 多重PCR 

分 类 号：S852.659.6[农业科学—基础兽医学] S852.659.1[农业科学—兽医学]