论文部分内容阅读
根据南方菜豆花叶病毒外壳蛋白基因序列设计并合成了4组RT-LAMP引物,通过引物筛选试验,确定SB1组引物为最佳引物,并进行了引物特异性与灵敏度检测试验,最终建立了南方菜豆花叶病毒的RT-LAMP检测方法。灵敏度检测试验显示,RT-LAMP方法比普通RT-PCR法灵敏度高10倍,而检测时间明显缩短,整个反应过程只需40 min。此外,体系中加入钙黄绿素,反应结束后,可裸眼观察颜色变化来判定结果。本研究所建立的SBMV RT-LAMP方法具有快速、稳定、灵敏、特异、操作简单的特点,适合于SBMV的现场快速检测。
Four sets of RT-LAMP primers were designed and synthesized based on the sequence of the coat protein gene of bean kidney mosaic virus in South China. Primer screening was used to determine the best primer for SB1 and the primer specificity and sensitivity were tested. Finally, RT-LAMP detection method for bean bean mosaic virus. Sensitivity test showed that the RT-LAMP method was 10 times more sensitive than the ordinary RT-PCR method, and the detection time was significantly shortened, the entire reaction process only 40 min. In addition, the system by adding calcein, after the reaction, the naked eye can observe the color change to determine the results. The method of SBMV RT-LAMP established in this study is fast, stable, sensitive, specific and easy to operate. It is suitable for on-site rapid detection of SBMV.