Quadruple fluorescent quantitative polymerase chain reaction (PCR) detection kit for porcine intestinal coronavirus

Abstract

The invention discloses a quadruple fluorescent quantitative polymerase chain reaction (PCR) detection kit for porcine intestinal coronavirus. The kit comprises eight six-base random reverse transcription primers having sequences shown in SEQ ID No. 1-SEQ ID No. 8, and four probe primers having sequences shown in SEQ ID No. 9-SEQ ID No. 12. The primers in the kit provided by the invention are specific primers which designed according to a highly conserved region of a conserved gene causing a main pathogen of porcine digestive tract virus diseases; a quadruple fluorescent quantitative PCR detection method is established for detecting porcine epidemic diarrhea virus (PEDV), porcine transmissible gastroenteritis virus (TEGV), porcine delta coronavirus (PDCoV) and porcine intestinal alpha coronavirus (PEAV); a most direct basis is provided for timely treatment, prevention and control of epidemic situations, and the sound development of the pig industry is escorted.

Description

technical field [0001] The invention relates to a veterinary detection kit and a method for using the detection kit in non-disease detection. Specifically, the invention is a 4-fold fluorescence quantitative PCR for porcine enteric coronavirus and a corresponding method for use. The above-mentioned 4-fold porcine viral diarrhea virus refers to: porcine epidemic diarrhea virus, porcine transmissible gastroenteritis virus, porcine delta coronavirus and porcine intestinal alpha coronavirus. Background technique [0002] Porcine viral diarrhea is one of the important infectious diseases that seriously harm the pig industry. The main traditional pathogens causing porcine viral diarrhea are porcine epidemic diarrhea virus (PEDV) and porcine transmissible gastroenteritis virus (TGEV). In recent years, porcine deltacoronavirus (PDCoV) and porcine enteric alphacoronavirus (PEAV) have been newly discovered, and these two coronaviruses are also seriously harmful to piglets. These viru...

AI Technical Summary

Problems solved by technology

These viruses are currently the main pathogens that cause viral diarrhea in pig herds or pig farms. They are very harmful, and there are mixed infections. The lethality rate of piglets ranges from 50% to 100%, causing great economic losses to the pig industry.

In addition, the clinical symptoms, pathological changes, and epidemic characteristics of viral diarrhea caused by the above-mentioned viruses are very similar. It is difficult to make a differential diagnosis based on clinical diagnosis alone, which often leads to misdiagnosis. Therefore, the prevention and control of the disease and its early diagnosis and differential diagnosis are particularly important

However, the complex etiology of diarrheal diseases and the limitations of existing diagnostic methods make it difficult for grassroots veterinarians and farms to diagnose diarrheal diseases, making the prevention and control of diarrheal diseases difficult to target and prescribe the right medicine

The most fundamental reason for this phenomenon is the lack of applicable fast, accurate and convenient commercial detection methods

For a long time, the detection kits for porcine viral diarrhea are mainly based on the traditional serological detection methods of porcine transmissible gastroenteritis virus (TGEV) and porcine epidemic diarrhea virus (PEDV). Two major changes occurred in the diarrhea epidemic: first, it mainly caused disease and death of suckling piglets, whose immune system was not yet fully developed, and failed to effectively produce serum antibodies; second, a highly pathogenic Virulent porcine deltacoronavirus and porcine enteric alphacoronavirus, there are no mature detection methods for these two viruses

It can be seen that the existing detection kits for detecting porcine viral diarrhea cannot meet the needs of current disease diagnosis, so there is an urgent need to develop a commercial detection kit that meets the current epidemic trend, and there is currently no one-time detection, diagnosis, Commercial kits for the identification and quantitative detection of the above four pathogens

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