非洲猪瘟病毒环介导恒温扩增快速检测技术的建立及应用  

DEVELOPMENT OF LOOP-MEDIATED ISOTHERMAL AMPLIFICATION(LAMP) FOR RAPID DETECTION OF AFRICAN SWINE FEVER VIRUS

作　　者：杨吉飞[1] 关贵全[1] 刘志杰[1] 汪月凤[1] 李有全[1] 马米玲[1] 刘爱红[1] 任巧云[1] 苟惠天[1] 杜鹏飞[1] 罗建勋[1] 殷宏[1] 

机构地区：[1]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甘肃省动物寄生虫病重点实验室农业部草食动物疫病重点实验室,兰州730046

出　　处：《中国动物传染病学报》2011年第4期7-12,共6页Chinese Journal of Animal Infectious Diseases

基　　金："948"项目(2010-S04);ASFRISK(211691);"973"项目(2010CB530206);国家自然科学基金项目(30800820;30972182;31072130;31001061);甘肃省重点项目(N1002NKDA035;0801NKDA033);农业部肉牛牦牛产业体系项目(CARS-38);科技部合作专项欧盟;欧盟EPIZONE(FOOD-CT-2006-016236);ARBOZOONET(211757);PIROVAC(KBBE-3-245145)资助

摘　　要：本研究基于非洲猪瘟病毒(African swine fever virus,ASFV)vp72基因设计引物,建立了能够快速检测非洲猪瘟病毒的环介导恒温扩增技术(loop-mediated isothermal amplification,LAMP)。将LAMP与OIE参考的PCR检测方法进行比较,并且应用LAMP对非洲猪瘟参考实验室提供的非洲猪瘟病毒17个毒株的基因组以及国内收集的50份猪的基因组、30份蜱的基因组进行检测。结果显示,本研究设计的引物具有良好的特异性和敏感性,所建立的LAMP能够成功扩增非洲猪瘟病毒17个毒株的基因组,而野外收集的猪和蜱的基因组检测均为阴性。因此,本研究所建立的方法能够用于非洲猪瘟的快速诊断以及防控。A loop-mediated isothermal amplification （LAMP） rapid assay was developed for detecting African swine fever virus （ASFV） based on the vp72 gene. This method was compared to the PCR which was recommended by OIE. Seventeen genomic DNAs of ASFV isolates from African swine fever reference laboratory, 50 field pig samples and 30 field tick samples from China were detected by LAMP, respectively. Seventeen genomic DNAs of ASFV isolates could be detected successfully, but all of the field samples were negative. The result revealed that this method had a good specificity and sensitivity, which could be used in rapid diagnosis, prevention and control of African swine fever.

关 键 词：环介导恒温扩增技术 聚合酶链式反应 非洲猪瘟 诊断 

分 类 号：S852.651[农业科学—基础兽医学] Q78[农业科学—兽医学]