番茄环斑病毒和烟草环斑病毒复合型胶体金免疫层析试纸条的研制  

Complex gold Immunochromatography assay for rapid detection of Tomato ringspot virus and Tobacco ringspot virus

作　　者：魏梅生[1] 杨翠云[2] 李桂芬[1] 

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

出　　处：《植物检疫》2008年第2期75-77,F0003,共4页Plant Quarantine

基　　金：国家质量监督检验检疫总局课题(2006IK216)

摘　　要：采用柠檬酸三钠还原法制备胶体金颗粒,标记番茄环斑病毒和烟草环斑病毒的兔多克隆抗体,用微定量喷头在硝酸纤维素膜上喷好2条病毒检测线(T线)和1条羊抗兔抗体质控线(C线),制成复合型免疫层析检测试纸条。一张试纸条在10min内,可同时检测出这两种病毒。试纸条检测番茄环斑病毒和烟草环斑病毒粗提纯液,检测灵敏度在1μg/mL,试纸条检测番茄环斑病毒和烟草环斑病毒的混合病汁液可稀释1000倍(重量/体积)。用健康叶片和缓冲液对照测试,试纸条结果都为阴性。Complex gold imrnunochromatography assay （GICA , Immunostrip） was developed by coupling colloidal golds with polyclonal rabbit antibody against Tomato ringspot virus or Tobacco ringspot virus. Two test lines and one control line on nitrocellulose filter were spread by Biojet Quanti 3000 dispenser on XYZ dispensing platform. The two viruses could be tested on one immunostrip within 10 minutes. The sensitivity of the complex immunostrip was 1 μ g/mL for the two partially purified viruses. Extract from ToRSV or TRSV infected Tetragonia expartsa or tobacco leaf was detected to a dilution of 10-3（W/V）. Non -specific reaction was not observed tor the healthy leaf and buffer control.

关 键 词：番茄环斑病毒 烟草环斑病毒 胶体金免疫层析法 免疫试纸条 

分 类 号：S432.4[农业科学—植物病理学] S41[农业科学—农业昆虫与害虫防治]