Dual fluorescence RT-PCR (Reverse Transcription-Polymerase Chain Reaction) detection method for PDCoV (porcine Delta coronavirus) and PEDV (porcine epidemic diarrhea virus) and application thereof

Abstract

The invention belongs to the field of microbiological detection, relates to the detection of a PDCoV (porcine Delta coronavirus) and a PEDV (porcine epidemic diarrhea virus), and particularly relates to a dual fluorescence RT-PCR (Reverse Transcription-Polymerase Chain Reaction) detection method for the PDCoV and the PEDV and application thereof. The detection method comprises the following steps of designing upstream and downstream primers of the PDCoV and upstream and downstream primers of the PEDV; preparing a dual real-time fluorescence system; implementing a dual real-time fluorescence detection method. The invention also provides the application of the dual fluorescence RT-PCR detection method to the simultaneous detection of the PDCoV and the PEDV. The detection method established by the invention can be used for simultaneously detecting the two viruses of the PEDV and the PDCoV quickly and specifically, and has favorable specificity, repeatability and sensitivity; the strong technical support is provided for the clinical diagnosis and prevention and treatment of the PEDV and the PDCoV; a foundation is also laid for the molecular epidemiological and prevention and control research of such epidemic diseases.

Description

technical field [0001] The invention belongs to the field of microbial detection and relates to the detection of porcine Delta coronavirus and porcine epidemic diarrhea virus, in particular to a double fluorescent RT-PCR detection method for porcine Delta coronavirus and porcine epidemic diarrhea virus and an application thereof. Background technique [0002] Porcine Deltacoronavirus (Porcine Deltacoronavirus, PDCoV) is a new type of porcine coronavirus, a member of the Coronaviridae family and the genus Coronaviridae, and is a single-stranded positive-sense RNA virus with an envelope. In 2012, Hong Kong scholar Woo first reported that PDCoV was detected in pig feces, and the virus was also detected in other mammals and birds. As of early 2014, PDCoV was also detected in piglet diarrhea in Ohio, USA. The clinical symptoms of PDCoV-affected pigs were severe diarrhea, vomiting, dehydration, and high lethality in piglets. Necropsy changes showed damage to the epithelial cells ...

AI Technical Summary

Problems solved by technology

However, it cannot meet the rapid detection of clinical PEDV and PDCoV mixed infection

[0005] Both PDCoV and PEDV are currently the main pathogens that cause diarrhea in piglets. Both can cause diarrhea in piglets to varying degrees. They have the characteristics of fast transmission, high infection rate, and wide epidemic range in pigs, and they have caused huge economic losses to the pig industry every year. loss

In particular, the mixed infection of PEDV and PDCoV is relatively common. It is difficult to distinguish the two pathogens by observing the clinical symptoms and pathological changes, which has caused great difficulties in clinical detection and control of these two diseases.

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