Quintuple RT-PCR detection kit for porcine viral diarrhea viruses

Abstract

The invention discloses a quintuple RT-PCR detection method and kit for porcine epidemic diarrhea viruses, porcine transmissible gastroenteritis viruses, porcine rotaviruses, porcine sapoviruses and porcine kobuviruses. The kit comprises ten specificity amplification primers. In the using process of the kit, total RNA of a sample to be detected is subjected to reverse transcription to become cDNA through a 6-basic-group random primer, a detection reaction system in the kit is used for RCR amplification with the cDNA as a template, and whether the sample to be detected is infected with one kind of pathogens or is under mixed infection is determined according to different sizes of amplified PCR segments of different pathogens. On the condition of ensuring specificity and sensitivity, the kit has the advantages of being easy and quick to operate and lowering detection cost and labor intensity, and is suitable for field sample detection.

Description

technical field [0001] The invention relates to a detection kit and a method for using the detection kit in non-disease detection. Specifically, the invention is a 5-fold RT-PCR detection kit for porcine viral diarrhea virus and a corresponding method of use. The 5-fold porcine viral diarrhea virus refers to: porcine epidemic diarrhea virus, porcine transmissible gastroenteritis virus, porcine rotavirus, porcine saapporo virus and porcine crest virus. Background technique [0002] Porcine viral diarrhea is one of the important infectious diseases that seriously endanger the pig industry. The main pathogens causing porcine viral diarrhea are porcine transmissible gastroenteritis virus (TGEV), porcine epidemic diarrhea virus (PEDV), and porcine rotavirus. Virus (RV), etc. In recent years, Sapovirus, Rigivirus and Norovirus have also been detected in pigs with diarrhea and healthy pigs respectively. These viruses are the main pathogens that cause viral diarrhea in pig herds o...

AI Technical Summary

Problems solved by technology

These viruses are the main pathogens that cause viral diarrhea in pig herds or pig farms, causing great harm and causing great economic losses to the pig industry every year

In addition, the clinical symptoms, pathological changes, and epidemic characteristics of viral diarrhea caused by the above-mentioned viruses are very similar, and it is difficult to make a differential diagnosis based on clinical diagnosis alone, which often leads to misdiagnosis. 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 available fast, accurate and convenient commercial diagnostic kits

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), porcine epidemic diarrhea virus (PEDV) and porcine rotavirus (RV) However, two major changes have taken place in the epidemic of porcine viral diarrhea in recent years: first, it mainly causes disease and death in suckling piglets, and the immune system of suckling piglets has not yet fully developed and failed to effectively produce serum antibodies; second, in recent years, In the epidemic of porcine viral diarrhea in China, new viruses such as porcine Sapporo virus and crest virus exist from time to time, and 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 and PCR commercial kits for differential diagnosis of the above five pathogens

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