鲤春病毒血症病毒恒温实时荧光快速检测方法的建立  

Rapid detection of spring viraemia of carp virus by real-time fluorescence loop-mediated isothermal amplification

作　　者：陈进会[1] 余霞[1] 余华[2,3] 黄伟[1] 邱杨[1] 刘建丽[1] 李红梅 石磊 叶蕾 

机构地区：[1]东莞出入境检验检疫局,广东东莞523072 [2]四川出入境检验检疫局,四川成都610001 [3]四川农业大学,四川雅安625014 [4]广州迪澳生物科技有限公司,广东广州510012

出　　处：《中国兽医科学》2013年第11期1146-1151,共6页Chinese Veterinary Science

基　　金：2011年东莞市科技计划资助项目(2011108102047)

摘　　要：根据鲤春病毒血症病毒(SVCV)的保守序列glyG基因,设计了3套LAMP引物,以ESE-Quant tube scanner为检测平台,通过简单易行的LAMP扩增检测SVCV,LAMP扩增温度为63℃。结果显示,在引入环引物后,恒温操作下30min内可得出结果,第1套LAMP引物显示出优良的扩增效率和扩增特异性;并且建立的LAMP法有极佳的灵敏度,其检测限可达到每反应2.4pg。与传统LAMP和荧光定量仪两种方式作为LAMP检测平台相比,ESE-Quant tube scanner平台具有简单、便携,试验过程数据化和自动化的特点,适合SVCV的现场检测和大规模监控。Three sets of loop-mediated isothermal amplification（LAMP） primers, and one loop primer were designed based on the conserved glyG sequence of spring viraemia of carp virus（SVCV）. Specificity and sensitivity of the assay were evaluated using a ESE-Quant tube scanner system. The amplification tern prature was 63 ℃. The results demonstrated that after added the loop primer,the amplification could be finished within 30 rain. The real-time LAMP assay was able to specifically detect SVCV without amplifica- tions of other viral RNA. The detection limit was 2.4 pg per reaction. The results suggested that this real- time LAMP method might have a great potential as a field usable molecular tool for diagnosis of SVCV.

关 键 词：鲤春病毒血症病毒 恒温实时荧光 环介导等温扩增 ESE—Quant TUBE SCANNER 

分 类 号：S852.659.5[农业科学—基础兽医学]