TaqMan探针实时荧光定量PCR检测番茄黄化曲叶病毒  

Detection of Tomato yellow leaf curl virus by real-time PCR with TaqMan probe

作　　者：阮美颖[1] 叶青静[1] 王荣青[1] 周国治[1] 姚祝平[1] 李志邈[1] 万红建[1] 杨悦俭[1] 

机构地区：[1]浙江省农业科学院蔬菜研究所,浙江杭州310021

出　　处：《浙江农业学报》2012年第5期842-845,共4页Acta Agriculturae Zhejiangensis

基　　金：国家科技支撑计划(2012BAD02B02);国家大宗蔬菜产业技术体系项目(CARS-25-G-16);浙江省公益技术研究农业项目(2011C22007)

摘　　要：根据番茄黄化曲叶病毒(Tomato yellow leaf curl virus,TYLCV)外壳蛋白(CP)基因序列上的保守序列,设计特异性的引物和Taqman探针,建立荧光定量PCR方法。结果表明,试验建立的标准曲线循环阈值(Ct值)与模板浓度具有良好的线性关系,相关系数为0.9941;能检测到1 000个病毒拷贝,灵敏度比普通PCR高100倍;与番茄花叶病毒和黄瓜花叶病毒无交叉反应,特异性强、重复性佳,为TYLCV检测提供了一种特异、灵敏、快速的定量检测方法。A quantitative real-time PCR assay was established for detection of Tomato yellow leaf curl virus. The primers and probe were designed according to the CP gene sequence of Tomato yellow leaf curl virus. Results indica- ted that the standard curve showed the linear relationship between Ct （cycle threshold） and template concentration, and the regression coefficient of the quantitative curve was 0. 9941. The sensitivity of the assay was 1 000 plasmid copies and it was 100 fold sensitive than conventional PCR. The specificity of the assay was high and there Was no cross reactions with ToMV and CMV. The real-time PCR system we established is rapid, sensitive and steady to detect TYLCV.

关 键 词：番茄黄化曲叶病毒 实时荧光定量PCR TAQMAN探针 定量检测 

分 类 号：S436.412[农业科学—农业昆虫与害虫防治]