猪塞尼卡病毒荧光定量RT-PCR检测方法的建立及初步应用  

Establishment and primary application of real-time RT-PCR for the detection of porcine Senecavirus A

作　　者：刘健新[1] 查云峰 鲁荣 任旭皎 黎振标 李慧子 张彭涛 宁章勇[1] LIU Jian-xin;ZHA Yun-feng;LU Rong;REN Xu-jiao;LI Zhen-biao;LI Hui-zi;ZHANG Peng-tao;NING Zhang-yong(College of Veterinary Medicine,South China Agricultural University,Guangzhou 510642,China;Guangdong Center for Animal Disease Prevention and Control,Guangzhou 510230,China)

机构地区：[1]华南农业大学兽医学院,广东广州510642 [2]广东省动物疫病预防控制中心,广东广州510230

出　　处：《中国兽医科学》2019年第3期274-279,共6页Chinese Veterinary Science

基　　金：广东省科技计划项目(2016A020210079)

摘　　要：本研究根据猪塞尼卡病毒(SVA)VP1基因保守序列设计引物,通过优化反应条件,建立了实时荧光定量RT-PCR检测方法,并对其进行特异性、敏感性和重复性的评价。结果显示,该检测方法特异性好,灵敏度达10 copies/uL,变异系数均小于2.0%,操作简单,耗时短。利用建立的检测方法对124份临床样本进行检测,结果,阳性率为3.2%。上述结果表明,本研究建立的荧光定量RT-PCR方法可用于猪原发性水泡病临床样品的快速检测和流行病学调查,具有良好的应用前景。In this research,a real-time RT-PCR detection method was established by optimizing reaction conditions based on primers designed according to the VP1 gene of Senecavirus A(SVA)and the specificity,sensitivity and reproducibility of the method were also evaluated.Results showed that the method had good specificity and the minimum detection limit was 10 copies/μL.The coefficient of variation was less than 2.0%with the merits of simple operation and little time-consuming.The detection of 124 clinical samples showed that the positive rate was 3.2%by this method.In conclusion,the real-time RT-PCR developed in this study could be used for the rapid detection of clinical samples with porcine idiopathic vesicular disease(PIVD)and the epidemiological investigation of SVA,and it had a good application prospect.

关 键 词：塞尼卡病毒 荧光定量RT-PCR 检测 

分 类 号：S852.659.6[农业科学—基础兽医学]