Fluorescence quantitative RT-PCR detecting agent for bean pod mottle virus, preparation method and application thereof

Abstract

The invention discloses a fluorescent quantitative RT-PCR detection reagent for a bean pod mottle virus, a preparation method thereof and application of the fluorescent quantitative RT-PCR detection reagent for the bean pod mottle virus. The fluorescent quantitative RT-PCR detection reagent for the bean pod mottle virus is synthesized through elaborate design by selecting conservative CP encodinggene fragments between various strains taking special bean pod mottle 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 98 bp; and the sequences of the primers and the probe are as follows: the sequences of the primers are BPMV TF: 5'-ttgatggcacaggaggaaatt-3' and BPMV TR: 5'-acatgcattgctgtagcttgag-3', and the sequence of the probe is 5'-FAM-tgtcttctagtgcaagtga-MGB-3'. The fluorescent quantitative RT-PCR detection reagent for the bean pod mottle 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 BPMV 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 the bean pod mottle virus, in particular to a fluorescent quantitative RT-PCR detection reagent for the bean pod mottle virus, a preparation method and an application. Background technique [0002] Bean pod mottle virus (BPMV) has natural hosts such as soybeans, cowpeas, and kidney beans. It mainly damages the leaves and seeds of soybeans. It causes mottled symptoms around the hilum, seriously affecting the quality of soybeans and reducing the quality of soybeans. output. The yield loss of soybeans infected by BPMV ranges from 3% to 52%. If it is co-infected with soybean mosaic virus, it can cause a yield loss of up to 85%. In the United States, BPMV has posed a serious threat to the soybean industry. BPMV can be transmitted by beetles such as the bean beetle beetle. At present, the virus is mainly distributed in the United States, Canada, Brazil and other countries. There is no report o...

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Problems solved by technology

[0006] 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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