猪塞内卡病毒A、猪水疱病毒和口蹄疫病毒三重荧光定量PCR检测方法的建立与应用  

Establishment and Preliminary Application of SVA,SVDV,FMDV Triplex Fluorescence Quantitative PCR Detection Method

作　　者：苏博 吴志敏 张馨月[1] 赵涵 鲍迪 张添琪 张嘉钰 梁湘雨 孟凡茹[1] 王开[1] 胡桂学[1] SU Bo;WU Zhi-min;ZHANG Xin-yue;ZHAO Han;BAO Di;ZHANG Tian-qi;ZHANG Jia-yu;LIANG Xiang-yu;MENG Fan-ru;WANG Kai;HU Gui-xue(College of Veterinary Medicine,Jilin Agricultural University,Changchun,Jilin,130118,China)

机构地区：[1]吉林农业大学动物医学院,吉林长春130118

出　　处：《动物医学进展》2023年第9期12-18,共7页Progress In Veterinary Medicine

基　　金：吉林省重点研发计划项目(20200402055NC)。

摘　　要：为建立一种鉴别猪塞内卡病毒A(SVA)、猪水疱病毒(SVDV)、口蹄疫病毒(FMDV)的方法,针对3种病毒的基因保守区域设计引物和探针,建立了可同时检测上述3种病毒的Taq Man三重荧光定量PCR方法。该方法仅能特异性扩增SVA和FMDV病毒基因以及SVDV阳性质粒,与猪繁殖与呼吸综合征病毒、经典猪瘟病毒、猪流行性腹泻病毒、猪圆环病毒2型、伪狂犬病病毒的阳性样品cDNA或DNA无交叉反应,对3种病毒核酸的最低检出限均为10 copies/μL,组内和组间变异系数均小于4%。应用该方法对2019-2022年采集的吉林省部分地区267份病料和本实验室留存的30份猪血清样品进行检测,未检出SVA、FMDV以及SVDV。建立的三重荧光PCR方法有利于对3种病毒进行有效的流行病学调查,可给快速准确鉴别致猪水疱症状病毒性疾病提供诊断方法。To establish a method for the simultaneous detection of Senecavirus A(SVA),Swine vesicular disease virus(SVDV)and Foot-and-mouth disease virus(FMDV),primers and probes were designed according to the conserved regions of the three viruses,a Taq Man triplex fluorescence quantitative PCR method was established and used to detect some clinical samples.The method was able to specifically amplify only SVA and FMDV nucleic acids and SVDV positive plasmids,and there was no cross reaction with cDNA or DNA of positive samples of porcine reproductive and respiratory syndrome virus,classical swine fever virus,porcine epidemic diarrhea virus,porcine circovirus type 2,and pseudorabies virus.The sensitivity of the method was 10 copies/μL for all three viruses,and the intra-and inter-group coefficients of variation were less than 4%.The method was used to detect 267 clinical samples collected from some areas of Jilin province and 30 pig serum samples kept in our laboratory between 2019 and 2022 respectively,and none of the above viral nucleic acids was detected.This study not only facilitated the epidemiological investigation of the three viruses,but also provided one of the diagnostic methods for the rapid and accurate identification of viral diseases with vesicular symptoms of swine.

关 键 词：塞内卡病毒A 猪水疱病毒 口蹄疫病毒 三重荧光定量PCR 

分 类 号：S852.65[农业科学—基础兽医学]