经典猪瘟荧光PCR检测方法的建立与应用  

Establishment and Application of Classical Fluorescent PCR Method for Detection of Swine Fever

作　　者：谷孝玉 李艳玲 裴雅芳 伏显华 刘静 解云辉 周兆言 李润[1] 李辰龙 GU Xiaoyu;LI Yanling;PEI Yafang;FU Xianhua;LIU Jing;XIE Yunhui;ZHOU Zhaoyan;LI Run;LI Chenlong(Tangshan Aon Bioengineering Co.,Ltd,Tangshan Hebei 063000,China)

机构地区：[1]唐山怡安生物工程有限公司,河北唐山063000

出　　处：《现代畜牧科技》2023年第10期10-13,共4页Modern Animal Husbandry Science & Technology

基　　金：河北省省级科技计划资助(21322914D);河北省省级科技计划资助(20322903D);唐山市科技创业创新领军人才项目资助(21130246A)。

摘　　要：猪瘟(CSF)已被世界卫生组织确定为A类传染病,为减少猪瘟病毒(CSFV)带来的经济损失,新的有效的诊断技术的开发成为关键。根据CSFV的Zaozhuang-1毒株的5'UTR基因保守区域设计出特异性引物,建立了一种猪瘟病毒荧光PCR快速检测方法。结果表明,该方法在62℃条件下扩增40个循环,灵敏度最高,最低可检测限度为10拷贝/μL;且特异性良好,与牛病毒性腹泻病毒(BVDV)、猪圆环病毒2型(PCV2)、猪繁殖与呼吸综合征病毒(PRRSV)和猪流行性腹泻病毒(PEDV)均无特异性扩增。对临床样本进行检测,结果显示,荧光定量PCR检测结果合格率为100%。该方法灵敏度高、特异性强、重复性良好、符合率高,适用于经典猪瘟的快速检测。Classical swine fever(CSF)has been identified as A class A infectious disease by the World Health Organization(WHO).In order to reduce the economic loss caused by Classical swine fever virus(CSFV),the development of new and effective diagnostic techniques has become the key.Based on the 5'UTR gene conserved region of CSFV strain Zaozhuang-1,specific primers were designed to establish a rapid detection method of swine fever virus by fluorescent PCR.The results showed that 40 cycles were amplified at 62℃with the highest sensitivity and the lowest detectable limit of 10 copies/μL.It is found to be related to Bovine Viral Diarrhea Virus(BVDV),Porcine Circovirus Type 2 Virus(PCV2),Porcine Circovirus Type 2 virus,Porcine Reproductive And Respiratory Syndrome Virus(PRRSV)and Porcine Epidemic Diarrhea Virus(PEDV)had no specific amplification.The clinical samples were tested,and the results showed that the qualified rate of fluorescence quantitative PCR was 100%.The method has high sensitivity,high specificity,good repeatability and high coincidence rate,and is suitable for the rapid detection of classical swine fever.

关 键 词：猪瘟病毒 荧光PCR 扩增 

分 类 号：S858.28[农业科学—临床兽医学]