猪流行性腹泻病毒RT-PCR检测方法建立及初步应用研究  

Development and Preliminary Application of a RT-PCR for Detection of Porcine Epidemic Diarrhea Virus

作　　者：吴学敏[1,2] 陈如敬[1,2] 王隆柏[1,2] 车勇良[1,2] 刘玉涛[1,2] 庄向生[1,2] 严山[1,2] 周伦江[1,2] 

机构地区：[1]福建省农业科学院畜牧兽医研究所,福州350013 [2]福建省畜禽疫病防治工程技术研究中心,福州350013

出　　处：《中国农学通报》2012年第26期59-62,共4页Chinese Agricultural Science Bulletin

基　　金：福建省农科院青年基金"猪流行性腹泻病毒TaqMan荧光定量RT-PCR检测方法的建立和初步应用"(2011QC-17);福建省农科院青年基金"国家公益性行业(农业)科研专项"(NYHYZX07-034);福建省农业科学技术项目"福建省生态养猪配套技术的研究与应用"(2009-03)

摘　　要：为建立一种特异、敏感的PEDV的检测方法。根据GenBank中公布的猪流行性腹泻病毒CV777株的M基因序列,设计了一对特异性引物,扩增长度为680bp的片段。将PCR产物进行测序,与CV777株、SH5株M基因的同源性分别为99.2%和97.6%。试验表明:该方法具有较好的特异性和敏感性,并能检测到PEDV的RNA浓度下限为100pg/μL。从福建省的3个地区共采集58份哺乳仔猪腹泻样品,并应用此方法进行检测,其阳性检出率为87.9%。In order to establish specific and sensitive detection methods of porcine epidemic diarrhea virus（PEDV）.A pair of primer was designed to amplify a product about 680 bp,according to the sequence of the M gene sequence of PEDV-CV777 strain of PEDV in Genbank.The amplified product was sequenced.Then the homology comparisons among the obtained sequence and the M Gene of the CV777 and SH5 stains were performed.The results showed that it shared 99.2% and 97.6% homology with M Gene of the CV777 and SH5,respectively.This method could be used to test PEDV,and the minimum detectable concentration of the PEDV RNA was 100 pg/μL.The method has been used to detect 58 portion specimens of diarrheal swine from three areas named A,B,and C in Fujian Province,and the positive tested rate of PEDV was 87.9%.

关 键 词：猪流行性腹泻病毒 RT-PCR M基因 

分 类 号：S855.3[农业科学—临床兽医学]