基于烟草蚜传病毒病的黄瓜花叶病(CMV)和马铃薯Y病毒病(PVY) LAMP检测体系研究

Studies on LAMP Detection System of Cucumber Mosaic Virus (CMV) and Potato Virus Y (PVY) Transmitted by Tobacco Aphids Virus

本文对基于烟草蚜传病毒病的黄瓜花叶病(CMV)和马铃薯Y病毒病(PVY),在优化LAMP体系、LAMP反应的最佳体系、LAMP产物的检测、LAMP灵敏性检测、田间样品的检测等几个方面进行了研究。结果表明:LAMP体系Mg2+的最佳浓度为6 mM,dNTPs的最佳浓度为1.2 mM;LAMP的最佳反应条件为反应温度63℃、反应时间为40 min;CMV和PVY的阳性LAMP产物显示为绿色,阴性LAMP产物显示为橘黄色;检测方法的灵敏度在DNA水平上可达到10~3 ng/μL;田间采集样品随机检测表明,建立的LAMP检测体系准确可靠。

Based on cucumber mosaic virus (CMV) and potato virus (PVY) transmitted by tobacco aphid, the optimization of LAMP system, the optimum system of LAMP reaction, the detection of LAMP products, the sensitivity of LAMP and the detection of field samples were studied. The results showed that the optimum concentration of Mg2+ in LAMP system was 6 mM;the optimum concentration of dNTPs was 1.2 mM;the optimum reaction condition of LAMP system was 63˚C;the reaction time was 40 min;the positive products of CMV and PVY were green;the negative products of LAMP were orange;the sensitivity of LAMP assay was 10 - 3 ng/L at DNA level. The results of random sampling in field showed that the LAMP assay was accurate and reliable.