Method for quickly detecting Chinese papaya leaf curl virus by LAMP (loop mediated isothermal amplification)

Abstract

The invention provides a method for quickly detecting Chinese papaya leaf curl virus by LAMP (loop mediated isothermal amplification), belonging to the field of plant protection. The method comprises the following steps of: designing an LAMP primer (P-F3, P-B3, P-FIP (feline infectious peritonitis) and P-BIP (bit interleaved parity)) according to the nucleotide sequence of the Chinese papaya leaf curl virus; performing the LAMP reaction after total DNA (deoxyribonucleic acid) is extracted from a sample; and performing the electrophoresis or the fluorescent dye to a reaction product. The method is simple to operate, some special apparatuses such as a PCR apparatus and the like are avoided, and the problems of the conventional methods such as the biology, the serology and the like are overcome, so that the detection method is simple, fast, sensitive and economical.

Description

technical field [0001] The invention relates to a method for rapidly detecting Chinese papaya leaf curl virus, which belongs to the field of plant protection. Background technique [0002] Chinese papaya leaf curl virus (Papaya leaf curl china virus, PaLCCNV) is an important plant virus, belonging to the Geminiviridae Begomovirus genus, and its vector is Bemisia tabaci. The virus was first detected on papaya in Guangxi, my country, so it is called Chinese papaya leaf curl virus. In addition to harming papaya, the virus also harms tomato and other crops. Symptoms such as yellowing between leaf veins and plant dwarfing have a serious impact on tomato yield and quality. In 2007, the disease ravaged tomatoes on a large scale in Henan, and 2,000 solar greenhouses in Kaifeng County, Kaifeng City alone were devastated, causing direct economic losses of nearly 20 million yuan. In 2009, it was also detected on crops in Jiangsu and other places, showing that the virus also showed sig...

AI Technical Summary

Problems solved by technology

[0004] At present, there are two commonly used methods for the detection of Panax melon leaf curl virus in China. One is the molecular biological detection method, among which PCR is the most common, but PCR requires 30-35 cyclic reactions (more time-consuming) and Special instrument (PCR instrument); Another kind is serological detection method, common have ELISA and DIBA etc., and serological detection method is except that there is deficiency in sensitivity, and the specificity of its detection is also limited (kidney bean golden mosaic leaf Relatively high nucleotide homology between members of the virus genus)

The LAMP method has overcome the shortcomings of the current two types of methods well (short reaction time, no need for special instruments, high detection sensitivity, good specificity), and there is no method for using this method to detect Chinese papaya leaf curl virus.

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