印第安纳型水泡性口炎病毒实时荧光RT-LAMP检测方法的建立  

Real time fluorescence RT-LAMP method for detection of vesicular stomatitis virus-Indiana

作　　者：李嘉阳 李津 王志锋 梁琳[1] 崔尚金[1] 李刚[1,3] LI Jia-yang;LI Jin;WANG Zhi-feng;LIANG Lin;CUI Shang-jin;LI Gang(Institute of Animal Sciences,Chinese Academy of Agricultural Sciences,Beijing 100193,China;R&D Center of Beijing Minhai Biotechnology Co.,LTD.,Beijing 102600,China;Institute of Zoology,Guangdong Academy of Sciences,Guangzhou 510260,China)

机构地区：[1]中国农业科学院北京畜牧兽医研究所,北京100193 [2]北京民海生物科技有限公司研发中心,北京102600 [3]广东省科学院动物研究所,广东广州510260

出　　处：《中国兽医科学》2021年第4期429-434,共6页Chinese Veterinary Science

基　　金：“十三五”国家重点研发计划项目(2016YFD0501102);国家自然科学基金面上项目(31472203);广东省“银龄专项”项目(2020A1313030136)。

摘　　要：利用ESE-Quant tube scanner监测平台,建立了一种基于实时荧光RT-LAMP技术的印第安纳型水泡性口炎病毒(VSV-IN)快速检测方法。根据VSV-IN的G基因序列,设计6条特异性引物,在63℃条件下,进行核酸扩增,反应时间为45 min。在扩增前加入SYBRGreenⅠ染料作为反应的指示剂,以SYBRGreenⅠ染料的荧光强度作为结果判定标准。结果表明,建立的实时荧光RT-LAMP方法操作简单、反应快速、灵敏度高、特异性好,能快速检测VSV-IN,并可以区分新泽西型水泡性口炎病毒(VSV-NJ)。Using ESE-Quant tube scanner monitoring platform,a fast detection method of Indiana vesicular stomatitis virus(VSV-IN)based on real-time fluorescent RT-LAMP technology was established.According to the gene sequence of G gene of VSV-IN,six specific primers were designed and amplified at63℃.The reaction time was 45 minutes.SYBR Green I dye was added as the indicator before amplification,and the fluorescence intensity of SYBR Green I dye was used as the result criterion. The results showed that the real-time fluorescent RT-LAMP method was simple,rapid,sensitive and specific,and could detect the Indiana type vesicular stomatitis virus(VSV-IN) quickly and can distinguish the New Jersey type vesicular stomatitis virus(VSV-NJ).

关 键 词：印第安纳型水泡性口炎病毒 新泽西型水泡性口炎病毒 实时荧光RT-LAMP 检测 

分 类 号：S852.659.5[农业科学—基础兽医学]