A rapid detection method for chlorotic ring spot-associated virus in kiwifruit based on rt-lamp-lfd

Abstract

The invention discloses a primer and a detection method for detecting Actinidia chlorotic ringspot-associated virus (AcCRaV) based on RT-LAMP-LFD; the primers include inner primers: FIP and BIP and outer primers: F3 and B3. The detection method comprises the following steps: S1, extracting the RNA of the kiwi fruit sample to be detected; S2, reverse-transcribing the extracted kiwi fruit sample RNA to be detected into cDNA; S3, using the cDNA of the kiwi fruit sample to be detected as a template to perform isothermal amplification; S4, utilizing The LFD test strip detects the isothermal amplified product obtained in step S3 to determine whether the kiwifruit sample to be tested contains kiwifruit chlorotic ring spot-associated virus. The invention designs LAMP primers according to the nucleotide sequence of the coat protein of kiwifruit chlorotic ring spot-associated virus, realizes the rapid detection of kiwifruit chlorotic ring spot-related virus, and can quickly identify whether the virus is contained in kiwifruit plants or tissue culture seedlings , so as to achieve the purpose of detection, monitoring, early warning and take targeted measures to reduce the losses caused by diseases.

Description

technical field [0001] The invention belongs to the technical field of plant virus detection, in particular to a rapid detection method for detecting kiwifruit chlorotic ring spot-related viruses based on RT-LAMP-LFD. Background technique [0002] Kiwi fruit is one of the most important horticultural products in the world. Because its pulp is rich in vitamin C, minerals and dietary fiber, it is favored by the public. In 2017, the world's kiwifruit production has reached 4.03 million tons, but the infection of diseases will affect the yield and quality of kiwifruit, and even affect the healthy development of the kiwifruit industry. Viral disease is a disease with a long incubation period and huge hidden dangers in production. Actinidia chlorotic ringspot-associated virus (ACRaV) is a negative-sense single-stranded RNA virus belonging to the genus Rowan ringspot virus in Europe. Under natural conditions, the virus can infect kiwifruit, and the leaves show symptoms such as ch...

AI Technical Summary

Problems solved by technology

In 2017, the world's kiwifruit production has reached 4.03 million tons, but the infection of diseases will affect the yield and quality of kiwifruit, and even affect the healthy development of the kiwifruit industry

The readiness of the test results depends on the quality of the electron microscope, and advanced instruments and equipment are often expensive

At the same time, inspectors must master good electron microscopy techniques and knowledge of virology

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