Fluorescence quantitative PCR detection reagent for Asf virus and preparation method and use thereof

Abstract

The related bio-reagent is designed by: with conservative African swine fever virus VP73 gene fragment as target object, designing and composing carefully for large quantities of primers and probes, taking optimal pairing screen test on different conditions to obtain the most proper primer and probe. The fluorescence quantitative RT-PCR testing reagent includes one couple of specific primers and one specific fluorescent probe, and the amplification fragment length is 66bp. This invention is fit to fast testing for African swine fever.

Description

technical field [0001] The invention relates to a biological agent designed and synthesized by taking the gene fragment of African swine fever virus as a target, especially a reagent capable of detecting the African swine fever virus, and a preparation method and application of the reagent. Background technique [0002] African swine fever (ASF) 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 countries and regions where the disease occurs, the mortality rate can be as high as 100%. Under natural conditions, ASFV is lodged in Ornithodoros ticks and recessively infected African wild boars, warthogs, South African wild boars and giant wild boars. So far only pigs have been found to show clinical signs. [0003] After African swine fever was first discovered in Kenya in East Africa in 1909 in domestic pigs brought by European immi...

AI Technical Summary

Problems solved by technology

Among them, the sensitivity and specificity of red blood cell adsorption test and direct immunofluorescence test are not high, and the animal inoculation test is not safe. ELISA has great practicability in quarantine and diagnosis, and is sensitive, specific, and the results are highly consistent. characteristics, but the ELISA method can only detect antibodies and cannot directly detect viruses

The polymerase chain reaction (PCR) method for detecting ASFV has the characteristics of specificity, sensitivity, and rapidity, but it has disadvantages such as time-consuming, easy to pollute, electrophoresis detection after amplification, and a small number of samples for each detection.

So far, there has been no report on the rapid detection method of African swine fever virus and related detection reagents in China.

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