胶体金免疫层析试纸结合RT-PCR法快速检测烟草环斑病毒  

The Rapid Detection of Tobacco Ringspot Virus by Gold Immunochromatography Assay and RT-PCR

作　　者：杨翠云[1] 魏梅生[2] 于翠[1] 乔艳艳[1] 

机构地区：[1]上海出入境检验检疫局,上海200135 [2]中国检验检疫科学研究院动植物检疫研究所,北京100029

出　　处：《上海交通大学学报（农业科学版）》2008年第3期208-211,共4页Journal of Shanghai Jiaotong University(Agricultural Science)

基　　金：国家质量监督检验检疫总局课题(2006IK216);上海市科委项目(06DZ05029;03DZ19314)

摘　　要：利用烟草环斑病毒胶体金免疫层析检测试纸条,对烟草环斑病毒3个分离物进行了快速检测。结果表明,试纸条在10min内,可特异性检出烟草环斑病毒的3个分离物,对番茄环斑病毒、南芥菜花叶病毒、番茄黑环病毒和健康叶片无反应。剪下试纸条上显示的检测带,进行RT-PCR,能扩增到与预期大小相同的DNA条带。试纸条检测烟草环斑病毒粗提纯液,检测灵敏度在1μg·mL-1,试纸条检测烟草环斑病毒的病汁液,检测灵敏度在10-3(W/V),试纸条检测灵敏度与DAS-ELISA方法灵敏度基本相同。Three isolates of tobacco ringspot virus were rapidly detected with gold immunochromatography assay （GICA , Immunostrip）. The results indicated that the 3 isolates of TRSV could be tested specifically on immunostrip within 10 minutes and no-specific reaction was observed for the ToRSV, ArMV, TBRV and healthy leaves. RT-PCR amplification could produce a specific DNA band of the expected size with the reaction band cut on immunostrip. The sensitivity of the immunostrip was 1μg·mL^-1 for the partial purified viruses. Extract from TRSV- infected tobacco leaves was detected to a dilution of 10^-3（W/V）. The sensitivity of the immunostrip and DAS-ELISA was almost the same.

关 键 词：烟草环斑病毒 胶体金免疫层析 试纸条 RT-PCR 

分 类 号：S41-30[农业科学—植物保护]