应用纳米磁珠实时荧光定量PCR方法检测番茄黄化曲叶病毒  

Detection of Tomato yellow leaf curl virus by real-time quantitative PCR combining with magnetic nanoparticles

作　　者：赵晓丽[1] 周琦[1] 孙宁[2] 邓丛良[1] 

机构地区：[1]北京出入境检验检疫局植物实验室,北京100026 [2]东南大学,南京210096

出　　处：《植物病理学报》2014年第2期221-224,共4页Acta Phytopathologica Sinica

基　　金：公益性行业(质检)科研专项(201110035)

摘　　要：番茄黄化曲叶病毒（Tomato yellow leaf curl virus，TYLcV）为双生病毒科（Geminiviridae）菜豆金色花叶病毒属（Begomovirus）成员，于1964年在以色列首次发现，目前我国已有15个省（自治区、直辖市）发现该病毒,TYLCV主要通过烟粉虱传播。In order to develop a rapid, sensitive and specific qPCR assay for detection and quantification of Tomato yellow leaf curl virus （TYLCV）, a pair of primers and TaqMan probe were designed according to the conserved sequence Of known TYLCV isolates. Combining with MNP technique, a novel MNP-qPCR detection method was established and verified based on specificity, sensitivity and reproducibility tests. The results indica- ted that the Ct value of plotted standard curve showed good linear relationship （ R2 = 0. 9994） with the log of copy number of template. The established method showed a high specificity for TYLCV detection without crossing reaction with Tomato severe leaf curl virus and Tomato yellow leaf curl Sadinia virus, and was 10-fold more sensitive than routine PCR. Both coefficients of variation were less than 2%, indicating a good reproducibility. We have provided a novel method for detection of TYLCV in plant samples rapidly and quantitatively.

关 键 词：黄化曲叶病毒 PCR方法 荧光定量 番茄 菜豆金色花叶病毒属 检测 实时 磁珠 

分 类 号：S436.412.1[农业科学—农业昆虫与害虫防治]