基于猪伪狂犬病毒gE基因的PCR法快速诊断猪伪狂犬病的研究  

A study of PCR assay for detection of pseudorabies based on PRV gE gene

作　　者：杨雪[1] 朱玲[1] 史小红[1] 郭万柱[1] 

机构地区：[1]四川农业大学动物生物技术中心动物疫病与人类健康实验室,四川雅安625014

出　　处：《黑龙江畜牧兽医》2010年第12期10-13,共4页Heilongjiang Animal Science And veterinary Medicine

基　　金："十一五"国家科技支撑计划项目(2006BAD06A18);"长江学者和创新团队发展计划"创新团队项目(IRT0848)

摘　　要：为了建立一种快速检测猪伪狂犬病毒(PRV)的方法,试验采用PCR法,并以NCBI公布的PRV Min-A株(登录号为AY170318.1)的gE基因序列为参考序列设计1对特异性引物,进行PRV基因的扩增,并对该检测方法的特异性、敏感性、重复性进行验证。结果表明:该PCR检测方法扩增的目的基因长348 bp;应用此方法对猪繁殖与呼吸综合征病毒(PRRSV)、猪圆环病毒Ⅱ型(PCV2)、猪瘟病毒(CSFV)、猪细小病毒(PPV)进行PCR扩增均未有条带出现;该方法能够检测到的最低DNA模板量为10 pg;重复性良好;应用此法对43份临床样品进行检测,检出率为65.12%(28/43)。说明该PCR检测方法可用于PRV的分离鉴定、临床病料检测和分子流行病学调查等。To establish a method for rapid diagnosis of pseudorabies,the experiment designed a pair of specific primers based on the gE gene sequence of PRV Min-A strains（accession number： AY170318.1） which was published in the NCBI PRV gene was amplified by using PCR technology,testing the PCR detection method established by specificity,sensibility,reproducibility.The results indicated that the size of target gene was 348 bp by PCR amplification.Negative test results were found on the porcine reproductive and respiratory syndrome virus,porcine circovirus type 2,classical swine fever virus and porcine parvovirus.As little as 10 pg of PRV DNA could be detected with high reproducibility.The PCR method were tested by 43 clinic suspicious samples,and 65.12%（28/43） of the samples were pointed out to be positive.This illustrateds that the PCR method can applied for the clinical investigation and epidemiological survey of brucellosis.

关 键 词：伪狂犬病毒 GE基因 PCR 检测 

分 类 号：S858.28[农业科学—临床兽医学]