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A kind of bovine coronavirus ELISA detection kit

A detection kit, coronavirus technology, applied in the direction of viruses, viral peptides, viruses/phages, etc., can solve the problems of high diagnostic cost, inability to use rapid diagnosis, and long diagnostic cycle.

Active Publication Date: 2020-09-04
HEILONGJIANG BAYI AGRICULTURAL UNIVERSITY
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Problems solved by technology

[0003] Generally speaking, there are many detection methods for bovine coronavirus, including general methods such as EM, IEM, IF, IFA, HA, RT-PCR, RTFQ PCR, ELISA and many other methods, although the first few methods can accurately detect BCoV Diagnosis, but these methods have a long diagnostic cycle and high diagnostic costs, and cannot be used for rapid diagnosis of large-scale cattle farms. When a large-scale disease outbreak occurs, accurate diagnostic results and shortened diagnostic time can be better. Reduce the economic loss of the cattle farm

Method used

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  • A kind of bovine coronavirus ELISA detection kit
  • A kind of bovine coronavirus ELISA detection kit
  • A kind of bovine coronavirus ELISA detection kit

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Embodiment

[0038] A bovine coronavirus ELISA detection kit, the kit contains recombinant N protein.

[0039] The recombinant N protein is the recombinant pET-32a-N protein of BCoV-CD isolate.

[0040] The standard positive serum and negative serum of BCoV were prepared and preserved by the Infectious Disease Research Laboratory of Bayi Agricultural University, Heilongjiang. The following pMD18-T was purchased from Dalian Bao Biological Engineering Co., Ltd.; the prokaryotic expression vector pET-32a and Escherichia coli BL21 were both preserved by the Infectious Disease Prevention Laboratory of Heilongjiang Bayi Agricultural University.

[0041] The recombinant N protein is a hydrophobic region with better immunogenicity predicted by DNAStar software.

[0042] The preparation method of the recombinant N protein comprises the following steps:

[0043] A primer design and synthesis

[0044] According to the isolated strain of BCoV-Mebus released on GenBank, the N gene primer sequence wa...

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Abstract

The invention discloses an ELISA (Enzyme-Linked Immuno Sorbent Assay) detection kit for bovine coronavirus. The ELISA detection kit is characterized in that purified BCoV-CD isolate recombinant pET-32a-N protein is used as coating antigen, reaction conditions are optimized, and an indirect ELISA diagnosis method for detecting bovine-serum characteristic N protein antibodies. The ELISA detection kit disclosed by the invention has the beneficial effects that escherichia coli is adopted for prokaryotic expression, the raw-material sources are wide, the preparation is easy, the standardization iseasy, and the production and detection processes are safer and more reliable, so that the ELISA detection kit is suitable for being promoted and used in basic units.

Description

technical field [0001] The invention relates to the technical field of virus detection, in particular to a bovine coronavirus ELISA detection kit. Background technique [0002] Diarrhea is one of the most serious diseases of newborn calves. Calves are at the greatest risk of developing diarrhea within the first month of life. The incidence of diarrhea decreases with age. Viruses, bovine coronavirus and enterotoxigenic Escherichia coli (K99) are related to four major enteric pathogens. Bovine coronavirus is the most important pathogen causing adult bovine winter dysentery (WD) and calf coronavirus diarrhea (CD). Through the investigation of antibody detection in animal serum, it was found that there is a high positive rate of bovine coronavirus in the three northeastern provinces. [0003] Generally speaking, there are many detection methods for bovine coronavirus, including general methods such as EM, IEM, IF, IFA, HA, RT-PCR, RTFQ PCR, ELISA and many other methods, althou...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/569C07K14/165C12N15/50C12N15/70
CPCC07K14/005C12N15/70C12N2770/20022G01N33/56983G01N2469/20
Inventor 周玉龙范春玲倪宏波朱战波侯喜林邵红武瑞任亚超高国强贾伟强王梦心胡林杰孟野盛守志王訢丞白彪慧黄晓毅
Owner HEILONGJIANG BAYI AGRICULTURAL UNIVERSITY
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