Rapid porcine delta coronavirus detection primer and kit as well as application thereof

Abstract

The invention provides a rapid porcine delta coronavirus detection primer and kit as well as an application thereof. A dual priming oligonucleotide (DPO) primer design technology is introduced, a DPOprimer is designed according to a highly conserved specific sequence of a porcine delta coronavirus (PDCoV), and a reaction system is optimized, so that a detection method for the PDCoV and the corresponding detection kit are established. The kit has the characteristics of rapidness, simplicity, convenience, strong specificity, high sensitivity and good reliability, can realize accurate detectionof the PDCoV, assists in prevention, control, diagnosis and treatment of the PDCoV, and has a good application prospect.

Description

technical field [0001] The invention belongs to the technical field of animal disease detection, and in particular relates to a rapid detection primer, kit and application of porcine delta coronavirus. Background technique [0002] Porcine delta coronavirus (PDCoV) was discovered through epidemiological investigation in 2012, and then PDCoV was isolated and identified on LLC-PK cells. PDCoV is a novel porcine coronavirus that can cause acute enteritis in neonatal piglets. So far, the United States, Canada, China, South Korea, Thailand, Japan and other countries have successively reported the existence of PDCoV, causing significant economic losses to the pig industry. [0003] PDCoV belongs to the Coronavirus family (Coronavirinae) and is a single-stranded positive-sense RNA virus with an envelope. It is the smallest coronavirus known today. The translated region (UTR), six open reading frames (ORF1a, ORF1b and ORF2 to 5) and a 3'UTR. This virus and other swine-derived cor...

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Problems solved by technology

This virus and other swine-derived coronaviruses can cause symptoms such as vomiting and diarrhea in pigs clinically, and it is difficult to distinguish them from clinical features and pathogenic forms

[0004] At present, domestic laboratory diagnosis methods include pathological examination, serological examination, and RT-PCR examination, etc. However, the existing detection methods have limitations and are easily limited by factors such as detection conditions, cycle, and accuracy, making it difficult to achieve efficient and rapid pathogenic diagnosis , such as virus isolation and serological detection methods are time-consuming, labor-intensive, low-sensitivity and prone to false positives

In recent years, the focus of PDCoV detection research is mainly on the PCR method. This method is widely used because of its strong practicability, high efficiency, rapidity and accuracy. However, the design process of conventional PCR primers is complicated, and repeated optimization of primer parameters is required. Poor, sometimes the occurrence of non-specific amplification cannot be avoided even if the reaction conditions are optimized

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