Triple fluorescent quantitative PCR detection primers and kit for identifying African swine fever wild strain and gene deletion strain

Abstract

The invention discloses triple fluorescent quantitative PCR (Polymerase Chain Reaction) detection primers and a kit for identifying an African swine fever virus wild strain and a gene deletion strain. The invention provides a group of specific primers and probes for detecting the African swine fever virus wild strain and the gene deletion strain, and the specific primers and probes are used for simultaneously detecting the B646L gene of the African swine fever virus wild strain and the EGFP and mCherry genes of the deleted strain through an RT-qPCR method, so that whether a sample is the African swine fever virus wild strain or the gene deletion strain can be identified. The detection method is high in sensitivity and strong in specificity, and the lowest detection concentrations of B646L, EGFP and mCherry gene positive standard plasmids are 5.94*10 <-9> ng / mu L, 9.67*10 <-9> ng / mu L and 5.85*10 <-9> ng / mu L respectively. The method is wide in detection range, can be used for detecting various African swine fever virus gene deletion strains, is rapid and efficient in detection process, is suitable for large-batch detection, provides a new means for identifying African swine fever virus wild strains and gene deletion strains, and can be used for rapidly identifying the African swine fever virus wild strains and gene deletion strains in production practice.

Description

technical field [0001] The invention relates to the technical field of biological detection, more specifically, to a triple fluorescent quantitative PCR detection primer and a kit for identifying African swine fever wild strains and gene deletion strains. Background technique [0002] African swine fever virus (ASFV) is the causative agent of a highly contagious and highly lethal domestic swine disease caused by the virus. Diseases have caused huge economic losses to the pig farming industry. At present, biosecurity and vaccines are the most effective and economical methods to prevent and control the ASF epidemic. Monitoring whether pigs are infected by pathogens is the top priority of biosecurity. [0003] ASFV gene deletion attenuated vaccine has become the most promising ASF vaccine candidate, and major swine disease research institutions in the world are vigorously developing effective ASFV gene deletion vaccines. At present, various research institutes and universitie...

AI Technical Summary

Problems solved by technology

[0004] At present, in the context of vaccine research on multiple gene-deleted strains, although patents CN111676327A, CN111074000A, and CN111020062A all disclose a real-time fluorescent quantitative PCR detection method and kit for distinguishing between African swine fever wild strains and gene-deleted strains, However, they are only limited to the detection of one or two gene deletion strains, and their application is obviously limited. Therefore, it is urgent to establish a new fluorescence quantitative method for more extensive identification of African swine fever wild strains and multiple mutant strains. PCR method

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