Fluorescent quantitative PCR detection reagent, kit and detection method for African swine fever virus

Abstract

The invention discloses a fluorescent quantitative PCR (Polymerase Chain Reaction) detection reagent, a kit and a detection method for an African swine fever virus (ASFV). The detection reagent is targeted to a conserved segment of an ASFV VP72 gene, and comprises a pair of specific primers, a specific probe and an internal labeling probe. The kit comprises a PCR mixed solution, Taq DNA polymerase, DEPC (diethylpyrocarbonate)-H2O, an ASFV positive quality product, an ASFV negative quality product, a quantification standard substance and a pseudovirus internal labeling solution, wherein the PCR mixed solution comprises a pair of specific primers, a specific probe and an internal labeling probe. The detection method comprises the steps of extraction of total DNAs, preparation of reaction components, making of a standard curve by diluting the standard substance, amplifying and result judgment. The reagent and the kit have the advantages of high specificity, high sensitivity, no pollution and high flux, and can be used for accurately detecting infection, inapparent infection or continuous carrying of a toxic host of low-content ASFV. The detection method is quick, specific and sensitive, and can be used for quantitatively detecting a large quantity of samples at the same time.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a PCR detection reagent, a kit and a detection method thereof, in particular to a fluorescence quantitative PCR detection reagent, a kit and a detection method of African swine fever virus. Background technique [0002] African swine fever virus disease is a contagious viral disease of pigs that attacks macrophages and monocytes and is typically characterized by high mortality, reddened areas of the skin and severe bleeding in internal organs, in the first occurrence of the disease In some countries and regions, the mortality rate can be as high as 100%. Under natural conditions, ASFV is lodged in orthodromos ticks and in recessively infected African wild pigs, warthogs, South African wild pigs and wild boars. The International Veterinary Bureau has identified the disease as a Class A animal infectious disease, and my country has also identified it as a Class A infectious disease for ...

AI Technical Summary

Problems solved by technology

Among them, the sensitivity and specificity of erythrocyte adsorption test and direct immunofluorescence test are not high, and the animal inoculation test is not safe

The polymerase chain reaction method for detecting African swine fever virus has the characteristics of specificity, sensitivity, and rapidity, but it is time-consuming, easy to contaminate, electrophoresis detection is required after amplification, and the number of samples for each detection is small.

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