An enzymatic visualized oligonucleotide chip for simultaneous detection of four porcine diarrhea viruses and its application

Abstract

The invention provides an enzymatic visualized oligonucleotide chip for synchronously detecting four porcine diarrhea viruses, which comprises a solid phase carrier and oligonucleotide probes immobilized on the solid phase carrier, which are used to detect porcine epidemics respectively Diarrhea virus, porcine transmissible gastroenteritis virus, porcine group A rotavirus, and porcine deltacoronavirus. The present invention also provides a method for preparing the above-mentioned chip. The invention also provides a kit and a detection method for synchronous detection of four porcine diarrhea viruses. The invention combines enzymatic reaction and gene chip technology, can quickly detect and differentially diagnose four kinds of porcine diarrhea virus diseases, has good specificity, high accuracy, and the detection results are visible to the naked eye, and provides a method for the prevention and control of porcine viral diarrhea A new diagnostic technology; moreover, the preparation method of the chip of the present invention is simple and low in cost, and can be applied to a large number of detections of clinical epidemic diseases, providing technical support for ensuring the healthy development of my country's pig industry.

Description

technical field [0001] The invention belongs to the field of molecular biology, and in particular relates to an enzymatic visualized oligonucleotide chip for synchronous detection of four porcine diarrhea viruses and an application thereof. Background technique [0002] In recent years, the occurrence and prevalence of porcine infectious diarrhea disease have become increasingly serious, causing incalculable economic losses to the majority of farmers. Porcine epidemic diarrhea virus (PEDV), porcine transmissible gastroenteritis virus (TGEV), porcine group A rotavirus , GAR). The appearance characteristics of the diseases caused by these three types of viruses are basically similar after the occurrence, and they all manifest as vomiting, watery jet diarrhea and dehydration, especially in sucking piglets with severe disease and high mortality. Porcine deltacoronavirus (Porcine Deltacoronavirus, PDCoV) was first reported in pig farms in Hong Kong in 2012, and then reported in...

AI Technical Summary

Problems solved by technology

[0005] At present, the report on the detection of porcine diarrhea virus by visual gene chip technology is only seen in the previous research of our team Ma Rui et al., Optimization of experimental conditions and application of gold-labeled silver-stained visual chip technology for porcine viral diarrhea, Journal of Agricultural Biotechnology, 2016, 24:1233 -1242, this method can only detect three specific porcine diarrhea viruses; moreover, the visualization chip used uses a glass slide as a solid phase carrier, which needs to be hydrated during preparation. This method is not only costly, but also has complicated operation steps and consumes duration

Images