Dual real-time fluorescence RT-PCR (reverse transcription-polymerase chain reaction) kit for detecting and identifying wild strain and vaccine strain of CSFV (classical swine fever virus) in swine umbilical cord blood and application of dual real-time fluorescence RT-PCR kit

Abstract

The invention discloses a dual real-time fluorescence RT-PCR (reverse transcription-polymerase chain reaction) kit for detecting and identifying a wild strain and a vaccine strain of a CSFV (classical swine fever virus) in swine umbilical cord blood and an application of the dual real-time fluorescence RT-PCR kit. The kit comprises a pair of primers and two fluorescent probes, wherein the sequences of the pair of primers are shown as SEQ ID NO.1 and SEQ ID NO.2, and the sequences of the two fluorescent probes are shown as SEQ ID NO.3 and SEQ ID NO.4. The kit has the advantages of high specificity, sensitivity and accuracy and excellent repeatability, meanwhile, the wild strain and the vaccine strain of the CSFV can be identified and detected by detecting the same sample once, and the problem of genetic crossover of the CSFV with the bovine viral diarrhea virus and the border disease virus which belong to the same genus can be solved. The kit is applicable to fast identification and detection of the wild strain and the vaccine strain of the CSFV in scientific research and clinical detection, can be used for precisely evaluating and diagnosing CSFV carrying and expelling conditions of sows and latent infection conditions of piglets and can be further used for evaluating effects of CSFV vaccines.

Description

technical field [0001] The invention relates to the technical field of molecular diagnosis of animal pathogens, in particular to a dual real-time fluorescent PCR kit for detecting and distinguishing wild strains and vaccine strains of classical swine fever virus in pig umbilical cord blood and its application. Background technique [0002] Classical swine fever is a highly contagious and acute infectious disease of pigs caused by classical swine fever virus (CSFV). Sexual spotting, multiple hemorrhage, necrosis and infarction in internal organs, contagious and high fatality rate. In recent years, great changes have taken place in the prevalence and incidence characteristics of CSF in my country. Mild CSF or atypical CSF caused by low-virulence CSF wild strains or persistent infection are more common. The pathological features are also different from typical swine fever, which brings great troubles to veterinary clinical diagnosis. [0003] As a class of infectious disease, ...

AI Technical Summary

Problems solved by technology

However, there are still few commercial kits independently developed in China

At the same time, the ordinary PCR detection method has the defects of prone to false positives, long operation time, nested amplification, and high technical requirements for technical operation.

[0005] The above diagnostic methods for swine fever have their own shortcomings in terms of sensitivity, specificity and timeliness, and these methods cannot effectively distinguish between wild strain infection and vaccination.

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