PCR结合核酸斑点杂交检测猪伪狂犬病毒  

A COMBINED PCR WITH DOT BLOT HYBRIDIZATION FOR PSEUDORABIES VIRUS

作　　者：乌那尔汗.吉斯汗 任衍倍 孟凡峰[2] 田似报 符玉涓 张继红 李舵 美热木古丽 任娟 林汉亮 崔治中[2,3,4] 常爽 赵鹏[2,3,4] WUNAERHAN.Jisihan;REN Yan-bei;MENG Fan-feng;TIAN Si-bao;FU Yu-juan;ZHANG Ji-hong;LI Duo;MEIREMUGULI;REN Juan;LIN Han-liang;CUI Zhi-zhong;CHANG Shuang;ZHAO Peng(Xinjiang Animal Health Inspection Institute,Urumuqi,830000,China;College of Animal Science and Veterinary Medicine,ShandongAgricultural University,TaVan,271018,China;Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Controland Prevention,Taiwan,271018,China;Shandong Provincial Engineering Technology Research Center of Animal Disease Control andPrevention,Tai’an,271018,China)

机构地区：[1]新疆动物卫生监督所,乌鲁木齐830000 [2]山东农业大学动物科技学院,泰安271018 [3]山东省动物生物工程与疾病防治重点实验室,泰安271018 [4]山东省畜禽疫病防制工程技术研究中心,泰安271018

出　　处：《中国动物传染病学报》2019年第1期85-89,共5页Chinese Journal of Animal Infectious Diseases

基　　金：新疆少数民族科技人才特殊培养计划(201423131);山东省双一流建设项目(SYL2017YSTD11)

摘　　要：本研究基于猪伪狂犬病毒(Porcine pseudorabies virus,PRV)gE基因,建立了一种PCR结合核酸斑点杂交检测方法来鉴别野毒株和疫苗株,并利用这一方法对山东省部分地区的猪临床病料中PRV进行了检测。结果显示,建立的PCR结合核酸斑点杂交检测方法只与PRV野毒株反应,而与PRV gE基因缺失疫苗、猪瘟病毒、猪流行性腹泻病毒、猪繁殖与呼吸综合征病毒以及猪圆环病毒2型不反应。PCR结合核酸斑点杂交方法在53份病料中检出PRV阳性病料5份,阳性率为9.43%,高于单纯PCR的检出率(7.55%)。结果表明,本研究建立的PCR结合斑点杂交的方法特异性强,灵敏度高,适合PRV野毒的检测以及流行病学调查,可应用于PRV的鉴别诊断和净化。In the present study, a combined PCR method with nucleic acid dot blot hybridization was developed for differentiation of Pseudorabies virus (PRV) vaccine strains from wild viruses based on the conserved sequences of gE gene. The results showed that the the combined PCR method with nucleic acid dot blot hybridization reacted with PRV wild-type viruses but not with gE gene deleted vaccine strains, Classical swine fever vims (CSFV), Swine epidemic diarrhea vims (PEDV), Porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV2). Afterwards, 53 samples were collected from diseased pigs in different regions of Shandong province for PRV diagnosis. As a result,9.43%(5/53) of these samples were PRV positive by the combined PCR with nucleic acid dot blot hybridization, while 7.55%(4/53) samples were positive by PCR only. These results suggest that this combined PCR method with nucleic acid dot blot hybridization might be used for differential diagnosis and epidemiological investigation of PRV.

关 键 词：猪伪狂犬病毒 核酸斑点杂交 GE基因 鉴别 

分 类 号：S852.659.1[农业科学—基础兽医学]