Fluorescence quantitative RT-PCR detecting agent for peanut stunt virus, preparation method and application thereof

Abstract

The invention discloses a fluorescent quantitative RT-PCR detection reagent for a peanut stunt virus, a preparation method thereof and application of the fluorescent quantitative RT-PCR detection reagent for the peanut stunt virus. The fluorescent quantitative RT-PCR detection reagent for the peanut stunt virus is synthesized through elaborate design by selecting conservative CP encoding gene fragments between various strains taking special peanut stunt virus as target sequences, and comprises a pair of special primers and a special fluorescent probe, wherein the length of an amplified target fragment is 76 bp; and the sequences of the primers and the probe are as follows: the sequences of the primers are PSV TF: 5'-cagatgccatccctcga-3' and PSV TR: 5'-ccaacgaagtgtacgtgtacc-3', and the sequence of the probe is 5'-FAM-catccaacctttgtttctag-MGB-3'. The fluorescent quantitative RT-PCR detection reagent for the peanut stunt virus overcomes the defects of time consumption, easy pollution, requirement of electrophoretic detection after amplification and so on of general RT-PCR, can quickly and accurately perform qualitative and quantitative detection on PSV in a sample, and has the advantages of simplicity, easy operation, directviewing result, high sensitivity, good repeatability and so on.

Description

technical field [0001] The invention relates to a biological preparation for detecting peanut dwarf virus, in particular to a fluorescent quantitative RT-PCR detection reagent for peanut dwarf virus, a preparation method and application thereof. Background technique [0002] Peanut stunt virus (PSV) belongs to the genus Cucumovirus, and its virions are spherical with a diameter of about 30nm. The virus has a wide range of hosts. Under natural conditions, it can infect peanuts, kidney beans, soybeans, tobacco, alfalfa, clover, black locust and other crops. It can cause leaf chlorosis and develop into green and light green flowers and leaves. New growth The leaves are usually yellow when they are unfolded, but they can turn into normal green. The leaves are narrow and small, and the leaf margins sometimes appear wavy and twisted; the diseased plants have few and small pods, and sometimes they are deformed or cracked. Peanut dwarf virus is mainly transmitted in the field by ap...

AI Technical Summary

Problems solved by technology

[0005] The Tm value of ordinary TaqMan probes is 65-72°C, and the fragment length is 25-40 nt. Due to its high specificity, if the probe does not completely match the target gene sequence, the detection efficiency will be low or even no fluorescent signal will occur. problems such as

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