PCV-2、PCV-3、PCV-4三重PCR鉴别检测方法的建立与应用  

Development and Application of Triple PCR for Detection and Differentiation of PCV-2,PCV-3 and PCV-4

作　　者：王勤[1] 欧云文 汪洋[3] 潘琴 刘俐君 何博[1] WANG Qin;OU Yunwen;WANG Yang;PAN Qin;LIU Lijun;HE Bo(Dazhou Vocational and Technical College,Dazhou 635001,China;Animal Disease Prevention and Control Center of Kaijiang County,Dazhou 636250,China;State Key Laboratory of Veterinary Etiological Biology,Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730046,China;Animal Disease Prevention and Control Center of Dazhou,Dazhou 635000,China)

机构地区：[1]达州职业技术学院,达州635001 [2]开江县动物疫病预防控制中心,达州636250 [3]中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室,兰州730046 [4]达州市动物疫病预防控制中心,达州635000

出　　处：《中国动物传染病学报》2024年第4期60-66,共7页Chinese Journal of Animal Infectious Diseases

基　　金：四川省科技计划项目(2020JDRC0146);甘肃省科技计划项目(20YF3NA005)。

摘　　要：为了建立一种快速鉴别检测猪圆环病毒2型(PCV-2)、猪圆环病毒3型(PCV-3)、猪圆环病毒4型(PCV-4)的方法,试验设计合成PCV-2、PCV-3、PCV-4特异性鉴别检测引物,经一步法三重RT-PCR反应条件的优化,建立了PCV-2、PCV-3、PCV-4三重PCR鉴别检测方法。该方法对PCV-2、PCV-3、PCV-4、PCV-2/PCV-3/PCV-4可分别扩增出216、415、678 bp、216 bp/415 bp/678 bp的特异性条带,检测PRV、PPV、PRRSV、CSFV、PEDV、TGEV均为阴性,对PCV-2、PCV-3、PCV-4的最低检测量分别为1.0×102、1.0×10^(3)、1.0×10^(3)拷贝/μL,与PCV-2、PCV-3、PCV-4单项PCR方法的符合率均为100%,应用该方法检测85份临床病料样品,PCV-2、PCV-3、PCV-4阳性感染率分别为21.17%、14.12%、4.71%。说明建立的PCV-2、PCV-3、PCV-4三重PCR鉴别检测方法具有良好的特异性、敏感性、准确性和临床适用性,为PCV-2、PCV-3、PCV-4的临床鉴别诊断提供一种简便、快速的分子生物学检测技术。To develop a rapid method for detection and differentiation of PCV-2,PCV-3 and PCV-4,the specific primers of PCV-2,PCV-3 and PCV-4 were designed and synthesized,for a triple PCR method.The specific bands of 216,415,678,216 bp/415 bp/678 bp were amplified from PCV-2,PCV-3,PCV-4 and PCV-2/PCV-3/PCV-4.The triple PCR method did not amplify PRV,PPV,PRRSV,CSFV,PEDV and TGEV.The minimum detection limits were 1.0×102 copies/μL for PCV-2,1.0×10^(3) copies/μL for PCV-3 and 1.0×10^(3) copies/μL for PCV-4.The coincidence rates of the single PCR method for PCV-2,PCV-3 and PCV-4 were all 100%.Then,85 clinical samples were tested by this method and the positive rates of PCV-2,PCV-3 and PCV-4 were 21.17%,14.12%and 4.71%,respectively.These results showed that the developed triple PCR method for differential detection of PCV-2,PCV-3 and PCV-4 had good specificity,sensitivity,accuracy and clinical applicability,thus provided a simple and rapid molecular biological diagnostic technology for PCV-2,PCV-3 and PCV-4.

关 键 词：PCV-2 PCV-3 PCV-4 三重PCR 鉴别检测 

分 类 号：S858.28[农业科学—临床兽医学]