猪传染性胃肠炎及流行性腹泻病毒二重RT-PCR检测方法的建立  

Establishment of a Duplex RT-PCR for Porcine Transmissible Gastroenteritis Virus and Porcine Epidemic Diarrhea Virus

作　　者：贾锐[1] 叶佳欣[1] 俞向前[2] 叶承荣[2] 朱建国[1] 

机构地区：[1]上海交通大学农业与生物学院,上海市兽医生物技术重点实验室,上海200240 [2]上海市浦东新区动物疫病预防控制中心,上海201299

出　　处：《上海交通大学学报（农业科学版）》2014年第4期71-75,94,共6页Journal of Shanghai Jiaotong University(Agricultural Science)

基　　金：浦东新区科技发展基金创新资金(PKJ2012-N13)

摘　　要：根据GenBank中已登录的猪传染性胃肠炎病毒(TGEV)、猪流行性腹泻病毒(PEDV)的序列,利用Primer 5.0设计合成了2对特异性引物。用这2对引物对TGEV、PEDV cDNA模板首先进行单项PCR条件优化,然后采用正交试验设计优化多重RT-PCR反应条件,结果同时扩增到2条与实验设计相符的492bp(PEDV)和211bp(TGEV)特异性条带,建立了能够同时检测2种病毒混合感染的特异、灵敏的多重PCR检测方法,可检测出约11pg的PEDV和13pg的TGEV。Based on the sequences of porcine transmissible gastroenteritis virus, （TGEV）and porcine epidemic diarrhea virus（PEDV）in GenBank, two pairs of special primers were designed. The PCR reaction of PEDV or TGEV was optimized first. Design L9 （34） orthogonal experiment to find optimal reaction conditions. Under the optimized reaction conditions, products of 492 bp for PRDV and 211 bp for TGEV were simultaneously amplified in the duplex PCR. The results showed that the duplex PCR for the detection of PEDV and TGEV with high sensitivity and specificity was developed with minimal detectable amounts of PEDV and TGEV around 11 pg and 13 pg,respectively.

关 键 词：猪传染性胃肠炎病毒 猪流行性腹泻病毒 多重RT-PCR 检测 

分 类 号：S858.28[农业科学—临床兽医学]