烟草脉带花叶病毒试纸条的制作及田间病株的快速检测  

Study on Colloidal Gold Strip in Detecting Tobacco Vein Banding Mosaic Virus

作　　者：纪玲玲[1] 成巨龙 朱信宁 吴云锋[1] 张建新[3] 李蓓[1] 侯伟[1] 孙润红 

机构地区：[1]西北农林科技大学植保学院与陕西省农业分子生物学重点实验室,陕西杨凌712100 [2]陕西省烟草公司,西安710003 [3]河南师范大学生命科学学院,河南新乡453007 [4]河南上蔡县农技中心,河南上蔡463800

出　　处：《西北农业学报》2009年第1期180-183,共4页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：陕西烟草病毒源鉴定与可持续控制技术研究(KJ-38-2008);教育部长江学者和创新团队发展计划(No.200558);高等学校学科创新引智计划(B07049)资助

摘　　要：建立一种快速、特异、简便的烟草脉带花叶病毒检测方法。应用胶体金标记技术和免疫层析原理,在玻璃纤维膜、硝酸纤维膜的检测带和质控带上分别喷上胶体金与烟草脉带花叶病毒兔多克隆抗体的偶联物、烟草脉带花叶病毒兔多克隆抗体和羊抗兔IgG,研制出烟草脉带花叶病毒快速检测试纸条。病汁液稀释1000倍后仍可快速检出,3-10 min即可显示结果。对烟草上危害严重的4种病毒进行测试,均无非特异性反应。4℃干燥密封保存的试纸条有效期大于2个月。用试纸条对田间病株进行随机检测,结果与RT-PCR检测结果一致。To develop a rapid, specific and easy assay for testing tobacco vein banding mosaic virus （TVBMV）, based on the principle of colloidal gold labeling technique and immunochromatography assay, the coupling product from colloidal gold and rabbit polyclonal antibody against TVBMV was sprinkled on the glass fiber film; rabbit polyclonal antibody and sheep anti-rabbit IgG were sprinkled on the test line and control line of nitrocellulose membrane, colloidal gold strip was executed to detect TVBMV quickly. Extract from infected tobacco leaf was detected to a dilution of 10^-3 and the test time was only 3-10 minutes. Non specific reaction was not observed for testing other four main viruses infecting tobaccos. And the test strips could be stored at 4℃ for more than two months. The test results in the field by strips accorded with those by RT PCR.

关 键 词：胶体金免疫层析法 烟草脉带花叶病毒 试纸条 

分 类 号：S435.72[农业科学—农业昆虫与害虫防治]