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

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.