PCR primer set and kit for detection of Pepper vein mottle viruses (PVMV) and application of PCR primer set and kit

Abstract

The invention discloses a PCR primer set for detection of Pepper vein mottle viruses (PVMV). The PCR primer set includes three nuclear inclusion protein b gene (NIb) specific primers of the Pepper vein mottle viruses and two reference gene Actin specific primers of solanaceous plants, wherein the three nuclear inclusion protein b gene (NIb) specific primers of the Pepper vein mottle viruses are PVMV-F1, PVMV-F310 and PVMV-R824, and the two reference gene Actin specific primers of solanaceous plants are PeActin-F1 and PeActin-R1093. Detection of PCR amplification results is performed in 1.0%-1.5% of agarose, no expensive fluorescent quantitative PCR instruments are needed, and the primer set has the characteristics of a high speed, economy, high stability, high accuracy and high sensitivity.

Description

technical field [0001] The invention belongs to the technical field of crop virus detection, and in particular relates to a PCR primer set, a kit and an application thereof for detecting sweet pepper vein mottle virus (PVMV). Background technique [0002] Pepper vein mottle virus (PVMV) belongs to the genus Potyvirus. The virus genome is a positive-sense single-stranded RNA with a size of about 9.8 kb. The 5' end is covalently bound to the VPg protein, and the 3' end has a Poly(A) tail structure. The virus can infect at least 35 different species of Solanaceae by rubbing inoculation, and can also infect plants in the Chenopodiaceae, Amaranthaceae, Rutaceae, and Cucurbitaceae. The symptoms of different PVMV strains infecting different hosts are different. The virus can cause yellowing of pepper leaves, distortion of leaf veins, chlorosis between veins, and even smaller and deformed leaves and fruits, and cause a large amount of yield reduction. The PVMV virus originated in ...

AI Technical Summary

Problems solved by technology

[0005] The present invention aims at the defects of two common detection methods for PVMV detection in the prior art: one is that the sensitivity and specificity of the serological detection method are not high, and non-specificity is prone to occur and cause false positive results; the other is that the RT-PCR method only uses A pair of specific primers for PVMV detection cannot rule out false negatives, and even false positives, resulting in technical problems that the detection results are not accurate enough. A PCR primer set for accurate detection of PVMV is provided, which can be used for pepper pathogenic plants from different regions. PVMV nuclear inclusion body protein b (Nuclear Inclusion body'b'protein, NIb) coding gene was sequenced, and 3 specific primers were designed from the conserved region to form 2 pairs of primers, which can improve the specificity of PVMV detection and at the same time To eliminate the effect of false positive amplification, add a pair of specific primers designed for the actin gene (Actin) of solanaceous plants as an internal reference, which can detect the quality of cDNA synthesis and play a role in eliminating false negative amplification

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