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Method for detecting equine coronavirus antibody and special diagnosis kit thereof

A coronavirus and antibody technology, which is applied in the field of detection of equine coronavirus antibodies and special kits, can solve the problems of cumbersome means, low cost and technical requirements, and high cost

Inactive Publication Date: 2007-08-15
NANTONG ENTRY EXIT INSPECTION & QUARANTINE BUREAU OF THE PEOPLES REPUBLIC OF CHINA
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Problems solved by technology

[0004] Although the RT-PCR method for detecting equine coronavirus has high sensitivity and specificity, the method is cumbersome, the cost is high, and the technical requirements are also high. If the serum of the horse to be tested is tested, then relatively Said that the detection process will be relatively simple, and the cost and technical requirements will be low

Method used

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Embodiment Construction

[0027] (1) Expression of equine coronavirus nucleocapsid gene fragment in prokaryotic system

[0028] According to the ECV nucleocapsid gene sequence published by NCBI, specific primers were designed, and the ECV nucleocapsid gene fragment was amplified by PCR; the gene fragment was inserted into the expression vector pGEX-6p-1, and transformed into Escherichia coli DH5α, and constructed Recombinant plasmid pGEX-ECV-N. The pGEX-ECV-N plasmid was extracted and identified correctly by BamH I and EcoR I double enzyme digestion, transformed into Escherichia coli BL21, induced expression with IPTG, and analyzed by SDS-PAGE gel electrophoresis, the results proved that GST-ECV was successfully expressed in this study -N fusion protein. The prokaryotic expression product of the fusion protein was immunogenic when the mice were immunized by gel cutting.

[0029] (2) Expression of equine coronavirus nucleocapsid gene fragment in eukaryotic system

[0030] ECV-N was excised from the p...

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Abstract

This invention discloses one method and special agent case to test horse head virus antibody, which comprises the following steps: the horse head virus nuclear shell gene expression product of polyphenylacetylene standard board; reaction board adding the horse blood serum to be tested; adding enzyme label IgG single clone antibody, color development and testing for OD490nm. This invention applies real nuclear expression system to re-create bar virus infected insect cell and to get the virus expression.

Description

Technical field: [0001] The invention relates to a detection method and kit for animal infectious diseases. Background technique: [0002] Equine coronavirus (ECV) is a single-stranded positive-sense RNA virus. In 1975, more than 40 cases of infectious disease in newborn foals in the United States, characterized by watery diarrhea, fever and lymphoid tissue lesions, were finally proved to be caused by equine coronavirus (ECV) infection. In 1983, Huang JC et al. isolated ECV from a sick horse with severe diarrhea. Since then, ECV has entered the sight of people's research and made some research progress. [0003] Establishing a specific and rapid ECV diagnostic method is the premise of controlling and eradicating ECV. At present, Gu Bingquan and others have established an effective RT-PCR diagnostic method in the Australian laboratory, which can detect ECV pathogens in feces and disease materials, with high sensitivity and specificity. On this basis, by optimizing the cond...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/569G01N33/547C12Q1/68
CPCY02A50/30
Inventor 顾炳泉秦爱建冉俊祥李建陶宏锦王金龙张敬友孙谦
Owner NANTONG ENTRY EXIT INSPECTION & QUARANTINE BUREAU OF THE PEOPLES REPUBLIC OF CHINA
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