Multiplex PCR primer group for detecting porcine epidemic diarrhea virus, porcine transmissible gastroenteritis virus and porcine rotavirus simultaneously

Abstract

The invention discloses a multiplex PCR primer group for detecting porcine epidemic diarrhea virus, porcine transmissible gastroenteritis virus and porcine rotavirus simultaneously, and belongs to the field of virus detection. The primer group comprises three pairs of primers, wherein the primer sequences of the first pair of primers used for detecting the porcine epidemic diarrhea virus are respectively SEQ ID NO:1 and SEQ ID NO:2, the primer sequences of the second pair of primers used for detecting the porcine transmissible gastroenteritis virus are respectively SEQ ID NO:3 and SEQ ID NO:4, and the primer sequences of the third pair of primers used for detecting the porcine rotavirus are respectively SEQ ID NO:5 and SEQ ID NO:6. Through the application of the sequences of the primers, different strains of the porcine epidemic diarrhea virus, the porcine transmissible gastroenteritis virus and the porcine rotavirus can be detected simultaneously through the multiplex PCR method, and the detection results of the porcine epidemic diarrhea virus, the porcine transmissible gastroenteritis virus and the porcine rotavirus are masculine, while the defection results of other common pig-derived viruses are feminine, in conclusion, the primer group is strong in specificity, and good in repeatability; the PCR detection is carried out after the virus cDNA is subjected to gradient dilution, which shows that the sensitivity of the primers is high.

Description

technical field [0001] The invention relates to a set of primers for detecting viruses, in particular to a set of multiplex PCR primers for simultaneous detection of porcine epidemic diarrhea virus, porcine transmissible gastroenteritis virus and porcine rotavirus, belonging to the field of virus detection. Background technique [0002] Epidemic diarrhea virus (PEDV) is the causative agent of porcine epidemic diarrhea (PED). It mainly harms piglets. Sick suckling piglets have watery diarrhea and vomiting symptoms. Newborn piglets often suffer from severe dehydration and death after infection. Weaned pigs and fattening pigs have watery diarrhea for 4 to 6 days after illness, and their growth and development are affected after recovery. Influenced by weight loss in the late fattening period. Due to the short course of the disease and rapid spread, the disease spreads widely in our country and countries all over the world, causing serious economic losses every year. [0003] ...

AI Technical Summary

Problems solved by technology

This method is currently a relatively general method, which is characterized by accuracy and simple operation, but it takes a long time, and the result error is large, and the cost of manpower and material resources is large, which cannot meet the needs of rapid detection in practical applications.

Immunofluorescence detection method, through the combination of fluorescein-labeled secondary antibody, amplifies the signal, and observes and identifies it through fluorescence microscope, which improves the detection sensitivity to a certain extent, but in immunofluorescence method, the fluorescent antibody is often not stable enough, and the judgment is subjective It is highly resistant and prone to cross-reactions. At the same time, special equipment such as fluorescence microscopes are required, so there are many limitations in application.

ELISA is currently the most widely used immunological detection method. It can be used as a rapid detection method for qualitative or semi-quantitative detection. Indirect ELISA, double antibody sandwich ELISA, competition ELISA and blocking ELISA have been developed. The detection time is shortened to a certain extent, but special instruments (such as microplate readers) are still required, and the operation process is still complicated

Images