Multiple fluorescent quantitative primer and probe combination capable of simultaneously diagnosing three high-incidence pathogens of porcine diarrhea and application of multiple fluorescent quantitative primer and probe combination

Abstract

The invention provides a primer and probe combination for simultaneously detecting a porcine epidemic diarrhea virus (PEDV), a porcine transmissible gastroenteritis virus (TGEV) and a porcine delta coronavirus (PDCoV). The primer and probe combination comprises three groups of primers and probes respectively aiming at a PEDV N gene, a TGEV N gene and a PDCoV N gene. The combination further comprises primers and probes aiming at a 18S rRNA sequence as an internal standard in detection. The invention further provides a multiplex fluorescent quantitative PCR kit for differential diagnosis of porcine epidemic diarrhea pathogens caused by PEDV / TGEV / PDCoV. The multiplex fluorescent quantitative PCR kit comprises reaction liquid packaged by the kit, and the primers and the corresponding probe sets are packaged separately or in a mixed mode. Through design, optimization and screening of the primers and the probes, three high-incidence pathogens (PEDV, TGEV and PDCoV) of porcine diarrhea can be simultaneously identified and diagnosed in one reaction, amplification of specific genes of the three pathogens does not interfere with one another, specificity is high, sensitivity is high, repeatability is good, and the virus infection condition causing porcine diarrhea can be accurately diagnosed. The primer and probe combination has high application value and generates huge economic benefits.

Description

technical field [0001] The invention belongs to the fields of molecular biology and veterinary medicine, and in particular relates to a combination of multiple fluorescent quantitative primers and probes for diagnosing three highly pathogenic diarrheal diseases (PEDV, TGEV and PDCoV). Background technique [0002] Porcine epidemic diarrhea (PED) and porcine transmissible gastroenteritis (TGE) are the most common diarrheal diseases in pig farms. Porcine delta coronavirus disease (PDCoV) is a diarrheal disease that has only appeared in my country in recent years, but its clinical symptoms Very similar to the previous two. At present, relevant vaccines against diarrheal diseases cannot provide complete protection for pigs, and there is no specific drug available, which seriously reduces the rate of feed remuneration, increases drug use and labor costs, and often causes major economic losses for pig farms. PED, TGE, and PDCoV are highly contagious enteric diseases, and the corre...

AI Technical Summary

Problems solved by technology

The second is that during the detection process, due to improper operation by operators or equipment defects, aerosol pollution is likely to occur, resulting in false positive results, making it impossible to formulate the best prevention and control plan, and it is easy to miss the best prevention and control time

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