Dual fluorescent quantitation RT-PCR detection method for classical swine fever virus (CSFV) and bovine viral diarrhea virus (BVDV)

Abstract

The invention relates to a dual fluorescent quantitation RT-PCR detection method for the classical swine fever virus (CSFV) and the bovine viral diarrhea virus (BVDV) and belongs to the technical field of fluorescent quantitation RT-PCRs. According to the detection method, two groups of primers and probes are used, wherein the sequence of an amplification product of the first primer and probe group is solely consistent with a gene sequence conservation zone of the classical swine fever virus, and the sequence of an amplification product of the second primer and probe group is solely consistent with a gene sequence conservation zone of the BVDV. In addition, the CSFV probe P only appears in the amplification product consistent with the gene sequence conservation zone of the classical swine fever virus, the BVDV probe P only appears in the amplification product consistent with the gene sequence conservation zone of the bovine viral diarrhea virus, and each primer and probe group can eliminate interference of the other primer and probe group. In this way, by means of the detection method, the CSFV can be differentiated from the BVDV, and the classical swine fever virus and the bovine viral diarrhea virus can be detected at the same time.

Description

technical field [0001] The invention relates to a double fluorescent quantitative RT-PCR detection method for classical swine fever virus and bovine viral diarrhea virus, and belongs to the technical field of fluorescent quantitative RT-PCR. Background technique [0002] Classical swine fever virus (CSFV for short) and Bovine viral diarrhea virus (BVDV for short) both belong to the family Flaviviridae and the genus Pestivirus. The pig and sheep industry has caused huge economic losses. BVDV is widely distributed in the world, and animals such as goats, sheep, pigs, deer and wallabies are the hosts and sources of infection of BVDV. The nucleotide sequence of BVDV and CSFV has about 60% homology, and the amino acid has about 85% homology. The polyclonal antibody has serological cross-reaction to the two viruses. First, CSFV and BVDV infect pigs under natural conditions, and their clinical symptoms and pathological changes are similar; second, in the production of veterinary ...

AI Technical Summary

Problems solved by technology

OIE has announced the primer probe sequence for detecting BVDV, and the annotation cannot distinguish classical swine fever virus; CN103397107A discloses "bovine viral diarrhea virus fluorescence quantitative detection kit", but only detects BVDV separately

Clinical test results show that the existing CSFV or BVDV fluorescent quantitative RT-PCR kits cannot distinguish CSFV from BVDV

At present, the combined detection method for CSFV and BVDV infection has not been perfected, and the detection of porcine BVDV is still in the exploratory stage; there is no related reagent that can simultaneously detect CSFV and BVD

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