猪瘟病毒微滴式数字PCR方法的建立  

Development of Droplet Digital PCR Method for Detection of Classical Swine Fever Virus

作　　者：高晓龙 王英超 李月[2] 沈光年 刘晓冬 周德刚 梅力 冯小宇 GAO Xiaolong;WANG Yingchao;LI Yue;SHEN Guangnian;LIU Xiaodong;ZHOU Degang;MEI Li;FENG Xiaoyu(Beijing Center for Animal Disease Control and Prevention,Beijing 102600,China;Beijing Veterinary Drug Control Institution,Beijing 102629,China)

机构地区：[1]北京市动物疫病预防控制中心,北京102600 [2]北京市兽药监察所,北京102629

出　　处：《中国动物传染病学报》2021年第2期28-33,共6页Chinese Journal of Animal Infectious Diseases

基　　金：北京市科委重大专项首都食品质量安全保障课题(Z171100001317013)。

摘　　要：为建立猪瘟病毒微滴式数字PCR方法,实现猪瘟病毒定量检测。本研究根据猪瘟病毒5’UTR基因的保守序列设计了引物探针,对反应条件进行了优化,同时对该方法的灵敏性、特异性、重复性进行了评估,并对临床样品进行了检测。结果显示:本方法的最低检测下限为3.2copies/μL,未发现与其他病毒有交叉反应,样本重复的变异系数为3.9%,对临床样品的检测结果与实时荧光定量PCR(GB/T 27540-2011)的检测结果完全一致。本研究建立的猪瘟病毒微滴式数字PCR灵敏度高、特异性强、重复性好,适用于猪瘟病毒的临床检测。A droplet digital PCR method of Classical swine fever virus(CSFV)was developed in the present study for quantitative detection of CSFV.Specific primers and probes were designed according to the conserved sequence of 5'UTR gene of CSFV and optimized for their working concentrations.The sensitivity,specificity and repeatability of this method were evaluated for its detection of clinical samples.The results showed that this method had the lowest limit of detection at 3.2 copies/μL,no cross reaction with other viruses,the coefficient of variation of sample repetition at 3.9%,and the same detection rate as the real-time quantitative PCR(GB/T 27540-2011).The droplet digital PCR method developed in this study was suitable for the clinical detection of CSFV.

关 键 词：猪瘟病毒 微滴式数字PCR 检测方法 

分 类 号：S852.651[农业科学—基础兽医学]