Establishment and Application of a LAMP Method for the Rapid Detection of Sweet potato leaf curl virus
LI Hua-Wei, LIU Zhong-Hua, ZHANG Hong, XU Yong-Qing, LI Guo-Liang, LIN Zhao-Miao, XU Guo-Chun, QIU Yong-Xiang, JI Rong-Cang, LUO Wen-Bin, TANG Hao QIU, Si-Xin*
Crop Institute Research Centre, Fujian Academy of Agricultural Sciences, Fuzhou 350013, China, Scientific Observing and Experimental Station of Tuber and Root Crops in South China, Ministry of Agriculture, Fuzhou 350013, China
Abstract：Sweet potato leaf curl virus (SPLCV), is one of the most important diseases on sweet potato (Ipomoea batatas), that infection reduced the yields of the most sweet potato growing area. Rapid and accurate detection of SPLCV is essential for diagnosis, prevention and control of the disease. In this study, a set of primers for loop-mediated isothermal amplification (LAMP) detection was designed based on the coat protein gene (cp) selected from the sequencing results of SPLCV,design a set of LAMP specific primers, with SYBR Green Ⅰ for evaluation indicator, optimize the reaction conditions, a rapid and visualized LAMP method for detection of SPLCV was established, this method were completed under isothermal conditions at 65 ℃ for 1 h. Specificity test results showed that SPLCV could be specifically detected by this method, and there was no cross reaction with other DNA pathogens such as other sweet potato DNA virus, Tomato yellow mosaic leaf curl virus (TYLCV) and Tobacco leaf curl virus (TLCV), indicated that the method had good specificity. The sensitivity results showed that the minimum detection limit of this method was 1 pg/μL of the SPLCV genomic DNA, which was 10 times of ordinary PCR. It showed that this method had high sensitivity and good repeatability. The technique of LAMP of SPLCV established in this study had the advantages of rapid amplification, high efficiency, good specificity, simple operation, no need for special instruments, visual observation results with the naked eye, and is suitable for rapid detection of SPLCV samples in the field and grass-roots departments.
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