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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。
The specific primers and Taqman probes were designed according to the conserved sequences of CP gene of Tomato yellow leaf curl virus (TYLCV), and the fluorescence quantitative PCR method was established. The results showed that the standard curve established by the experiment had a good linear relationship between the Ct value and the template concentration, the correlation coefficient was 0.9941; 1 000 copies of the virus were detected with a sensitivity of 100 times higher than that of the normal PCR; And cucumber mosaic virus no cross-reaction, specificity and repeatability, provide a specific, sensitive and rapid quantitative detection method for TYLCV detection.