Triple fluorescent quantitative PCR kit for simultaneously detecting bovine rotavirus, bovine coronavirus and bovine viral diarrhea virus and application method of triple fluorescent quantitative PCR kit

Abstract

The invention provides a triple fluorescent quantitative PCR kit for simultaneously detecting a bovine rotavirus, a bovine coronavirus and a bovine viral diarrhea virus and an application, and belongs to the technical field of virus detection. The triple fluorescent quantitative PCR kit comprises three groups of specific primers and three corresponding probes aiming at the bovine rotavirus, the bovine coronavirus and the bovine viral diarrhea virus. The primer and the probe for detecting the three viruses have the advantages of strong specificity and high reaction efficiency. The triple real-time fluorescent quantitative PCR kit has the characteristics of high amplification efficiency, strong specificity, stable detection effects and good repeatability, and is suitable for dairy cow diarrhea pathogen detection, regular monitoring, dairy cow diarrhea epidemiological investigation and the like. The kit disclosed by the invention can be used for simultaneously detecting three dairy cow diarrhea pathogens in a real-time fluorescent quantitative PCR reaction system, so that the detection time is saved, the detection cost is reduced, a plurality of samples can be simultaneously detected, and the detection efficiency is high.

Description

technical field [0001] The invention relates to a bovine diarrhea pathogen detection kit, in particular to a kit for simultaneously detecting bovine rotavirus, bovine coronavirus and bovine viral diarrhea virus, and belongs to the technical field of virus detection. Background technique [0002] Bovine rotavirus, coronavirus and bovine viral diarrhea virus are important pathogens causing bovine viral diarrhea. With the large-scale and intensive development of my country's dairy farming industry, bovine diarrhea, especially neonatal calf diarrhea, has become one of the important diseases that plague the dairy farming industry, seriously endangering the health of the cow herd and hindering the development of the dairy farming industry in China. [0003] Bovine rotavirus (Bovine Rotavirus, BRV) disease is an acute gastrointestinal infectious disease caused by rotavirus in a variety of young animals and infants, characterized by mental fatigue, anorexia, vomiting, diarrhea, and ...

AI Technical Summary

Problems solved by technology

Among them, the detection techniques of nucleic acid hybridization and polyacrylamide gel electrophoresis are complex and costly, and are not suitable for clinical detection

RT-PCR, nano-PCR, fluorescent quantitative PCR and other detection technologies have good specificity and high detection sensitivity, and can be used for detection of tissue, blood, stool and other samples; however, since RT-PCR and nano-PCR are not suitable for batch detection, in production The application is still limited; at the same time, the RT-PCR method needs electrophoresis detection after detection, and the detection sensitivity is low. It is gradually replaced by real-time fluorescent quantitative PCR in rapid detection.

[0011] Fluorescent quantitative PCR method is a fast, accurate and high-throughput virus detection technology, which has been widely used in the rapid diagnosis of pathogens such as hepatitis C and AIDS in recent years; however, the existing methods for bovine rotavirus, coronavirus, or bovine viral diarrhea The virus detection methods are single-plex fluorescent quantitative PCR detection. In actual production applications, single-plex fluorescent quantitative PCR detection technology cannot meet the clinical needs of rapid, accurate, and high-throughput differential diagnosis of various viral pathogens that cause calf diarrhea.

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