PRV、PPV、PCV2和ASFV四重qPCR检测方法的建立及初步应用  

Establishment and preliminary application of quadruple qPCR method for PRV,PPV,PCV2 and ASFV

作　　者：陈旭 汤德元[1] 曾智勇[1] 王彬[1] 袁盛林 廖正波 何松 周飘 毛茵茗 CHEN Xu;TANG Deyuan;ZENG Zhiyong;WANG Bin;YUAN Shenglin;LIAO Zhengbo;HE Song;ZHOU Piao;MAO Yinming(College of Animal Science,Guizhou University,Guiyang 550025,China)

机构地区：[1]贵州大学动物科学学院,贵州贵阳550025

出　　处：《中国兽医学报》2025年第2期175-180,194,共7页Chinese Journal of Veterinary Science

基　　金：贵州省科技支撑计划资助项目(黔科合支撑[2021]一般162)。

摘　　要：为了鉴别临床上以繁殖障碍及流产为特征的病毒性疫病,本研究建立了同时检测猪伪狂犬病病毒(PRV)、猪细小病毒(PPV)、猪圆环病毒2型(PCV2)和非洲猪瘟病毒(ASFV)的四重qPCR方法,根据NCBI基因库中4种病毒保守基因设计4对特异性引物和探针,对反应的退火温度、引物浓度和探针浓度进行优化,并检验该方法的特异性、敏感性和重复性。结果显示:该方法不能检测到除目的病原以外的其他病原;对PRV、PPV、PCV2和ASFV 4种病原的最低检测限均为10拷贝;组内和组间重复性试验显示不同批次试验间C_(t)值变异系数均小于3%,说明该方法具有高度特异性、敏感性和稳定性。以上试验结果证明,本研究建立了高效灵敏的四重qPCR方法,为临床上猪伪狂犬病毒病、猪圆环病毒病、猪细小病毒病和非洲猪瘟的防控提供技术参考。To identify clinical viral diseases characterized by reproductive disorders and abortion,a quadruple qPCR method was established for simultaneous detection of PRV,PPV,PCV2 and AS-FV.Four pairs of specific primers and probes were designed according to the conserved genes of four viruses in the NCBI gene bank.The annealing temperature,primer concentration and probe concentration of the reaction were optimized,and the specificity,sensitivity and repeatability of the method were tested.The results showed that the method could not detect other pathogens except the target ones.The minimum detection limit of PRV,PPV,PCV2 and ASFV was 10 copies.Intra-group and inter-group repeatability tests showed that the coefficient of variation of C,values be-tween different batches was less than 3%,indicating that the method was highly specific,sensitive and stable.Establishment of an efficient and sensitive quadruple qPCR method provides technical reference for the clinical prevention and control of porcine pseudorabies virus disease,porcine circo-virus disease,porcine parvovirus disease and African swine fever.

关 键 词：猪伪狂犬病病毒 猪细小病毒 猪圆环病毒 非洲猪瘟病毒 四重qPCR 

分 类 号：S852.65[农业科学—基础兽医学]