一种基于分子信标荧光探针快速检测烟草花叶病毒的新方法  

A Novel Convenient and Fast Method for Detection of Tobacco Mosaic Virus-Ribonucleic Acid Based on Molecular Beacon

作　　者：刘凌凤[1] 王柯敏[1] 谭蔚泓[1] 李军[1] 孟祥贤[1] 郭秋平[1] 唐志文[1] 刘选明[1] 李杜[1] 

机构地区：[1]湖南大学生物技术研究院,化学化工学院,化学生物传感与计量学国家重点实验室,长沙410082

出　　处：《分析化学》2003年第9期1030-1035,共6页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金重点项目 (No .2 0 13 5 0 10 );海外青年学者合作研究基金 (No .2 0 0 2 85 0 6)资助课题

摘　　要：研究了一种利用新型荧光探针———分子信标进行植物病毒检测的方法 ,可以不要求对病毒RNA进行严格的分离和纯化 ,就能快捷地得到检测结果。此方法被用于烟草花叶病毒 (tobaccomosaicvirus,TMV)基因组RNA的检测 。A new approach for the detection of plant virus was developed with the use of a novel nucleic acid probe-molecular beacon. An antisense molecular beacon (MB1) was designed for the determination of the genome ribonucleia acid (RNA) of tobacco mosaic virus (TMV) with no need of stringent isolation and purification of the virus particles and RNA as well, which benefits from the prominent specificity and sensitivity of molecular beacon. Another sense molecular beacon (MB2) was also synthesized for control experiments. The result shows that the molecular beacon designed in consistent with the complementary sequence of a section of the conservative regions of TMV-RNA is suitable for the direct assay of TMV-RNA. Another group of control experiments has been performed to testify the experiment results more sufficiently and to achieve precise detection of TMV-RNA. Uninfected tobacco leaves were treated through the same process as that for infected leaves to get TMV-RNA. The obtained substance was used to react with the two MBs in the same situation as that for TMV-RNA. As a result, there are significant differences between the enhancement efficiencies of fluorescence intensity caused by the reactions of MB1 with the substances extracted from infected tobacco leaves and uninfected tobacco leaves. On the contrary, there's almost no differences for MB2. The result indicates that the proposed method is reliable for the direct detection of TMV-RNA.

关 键 词：分子信标 荧光探针 检测方法 烟草花叶病毒 植物病毒 分离 纯化 核糖核酸 

分 类 号：S435.72[农业科学—农业昆虫与害虫防治]