Porcine Delta coronavirus detection kit based on constant temperature isolation type fluorescent polymerase chain reaction (PCR) platform, and application of porcine Delta coronavirus detection kit

Abstract

The invention discloses a porcine Delta coronavirus detection kit based on a constant temperature isolation type fluorescent polymerase chain reaction (PCR) platform, and application of the porcine Delta coronavirus detection kit. The porcine Delta coronavirus detection kit consists of reaction buffer, a fluorescent quantitative PCR reaction liquid freeze-drying tube, a positive reference substance freeze-drying tube and a negative reference substance freeze-drying tube, wherein the fluorescent quantitative PCR reaction liquid freeze-drying tube is formed by mixing and freeze-drying Taq enzyme, reverse transcriptase, a detection primer, a probe and deoxynucleoside triphosphate (dNTP). The kit provided by the invention is short in detection time (the reaction time is only 42min), high in specificity and sensitivity and convenient to store (at 4 DEG C); when being matched with a constant temperature isolation type PCR instrument for use, the kit is very suitable for the rapid field detection of the porcine Delta coronavirus and can be widely popularized at the grassroots level.

Description

technical field

[0001] The invention belongs to the field of biotechnology, and in particular relates to a porcine Delta coronavirus detection kit based on a constant temperature isolation type fluorescent PCR platform and its application. Background technique

[0002] Porcine Deltacoronavirus (PDCoV) was first identified in molecular surveillance studies in Hong Kong in 2009, but its clinical significance was not investigated. The disease first appeared in Ohio in February 2014 and quickly spread to other states in the United States. PDCoV was also found in pigs in South Korea in April 2014. Recently, the disease has also appeared in mainland China and Thailand. Studies have shown that PDCoV can cause gastric alveolar epithelial necrosis, small intestinal villi atrophy, and mild interstitial lung. Animal experiments have confirmed that PDCoV can cause severe diarrhea, vomiting, and enteritis. The disease has a high incidence rate and a mortality rate of 30%-40%, which ha...

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