5种猪病毒性传染病病原的多重PCR检测方法的建立  

Development of multiplex PCR for detection of 5 pathogenic viruses to pigs

作　　者：冷依伊 任梅渗 蒙正群 张鹏飞[1,2] 杨泽晓[1] 姚学萍[1] 王印[1,2] LENG Yi-yi REN Mei-shen MENG Zheng-qun ZHANG Peng-fei YANG Ze-xiao YAO Xue-ping WANG Yin(College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China Key Laboratory of Animal Disease and Human Health of Sichuan Province, Chengdu 611130, China)

机构地区：[1]四川农业大学动物医学院,四川成都611130 [2]动物疫病与人类健康四川省重点实验室,四川成都611130

出　　处：《中国预防兽医学报》2017年第2期123-126,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家"十二五"国家科技支撑计划(2013BAD12B04)

摘　　要：为建立同时检测非洲猪瘟病毒(ASFV)、水疱性口炎病毒(VSV)、猪口蹄疫病毒(FMDV),猪瘟病毒(CSFV)以及猪伪狂犬病病毒(PRV)的多重RT-PCR检测方法,本研究根据Gen Bank中登录的参考病毒株序列,选择各病毒的保守序列设计5对特异性引物,通过优化反应条件,建立了一种可以同时快速检测以上5种病毒的多重RT-PCR检测方法。结果显示,该方法对不同的细菌或病毒模板扩增结果均为阴性,特异性强;敏感性试验表明该方法对PRV、CSFV、ASFV、VSV和FMDV的核酸最少检出量分别为8.82×10~3拷贝/μL、6.87×10~4拷贝/μL、5.71拷贝/μL、4.93×10~4拷贝/μL和4.32×10~2拷贝/μL。以上结果表明该方法快速、灵敏、特异性强,对以上5种猪病病毒能够进行快速鉴别检测,为其临床诊断与流行病学调查提供了有效的检测方法。In order to establish an rapid method to detect Africa swine fever virus （ASFV）, vesicular stomatitis virus （VSV）, foot and mouth disease virus （FMDV）, classical swine fever virus （CSFV） and porcine pseudorabies virus （PRV）. In this study, the method of multiplex PCR for detection of these 5 diseases was developed with the primers designed and synthesized according to the conservative sequences in reference strains of each kind of the virus. Through the optimization of reaction conditions, the method was specific for the 5 virus assay, but no any amplification was found for related pathogenic viruses. The detection limit of this method was 8.82x10^3, 6.87 x10^4, 5.71, 4.93 x10^4 and 4.32 x10^2 copies/μL for PRV, CSFV, ASFV, VSV and FMDV, respectively. These data indicated that the rapid, sensitive and specific detection method could be applied in these 5 virus detections for clinical diagnosis and pathogenic epidemiologic investigation.

关 键 词：多重PCR 非洲猪瘟病毒 猪口蹄疫病毒 猪水疱性口炎病毒 猪瘟病毒 猪伪狂犬病病毒 

分 类 号：S852.65[农业科学—基础兽医学]