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CCV (canine corona virus) M protein monoclonal antibody and preparation method thereof, and preparation method of immune colloidal gold test strip

A colloidal gold and monoclonal antibody technology, which is applied in the fields of biotechnology and animal disease quarantine and detection, can solve the problems of inconvenience for veterinary personnel to use clinically or in the field, and achieve the effects of simple method, broad application prospect and high sensitivity

Active Publication Date: 2019-07-05
YANGZHOU UNIV
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Problems solved by technology

However, these methods require special equipment and skilled professional technicians, and are not convenient for veterinary personnel to use in the clinic or in the field.

Method used

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  • CCV (canine corona virus) M protein monoclonal antibody and preparation method thereof, and preparation method of immune colloidal gold test strip
  • CCV (canine corona virus) M protein monoclonal antibody and preparation method thereof, and preparation method of immune colloidal gold test strip
  • CCV (canine corona virus) M protein monoclonal antibody and preparation method thereof, and preparation method of immune colloidal gold test strip

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Embodiment Construction

[0044] Source of biological material:

[0045] Canine coronavirus (JS1706, JS1712 strains) was isolated and identified by the Key Open Laboratory of Livestock and Poultry Infectious Diseases, Ministry of Agriculture, Yangzhou University.

[0046] Porcine transmissible gastroenteritis virus (TGEV), canine distemper virus (CDV), canine adenovirus (CAV-2), canine parvovirus (CPV-2a) virus strains, the focus of livestock and poultry infectious diseases of the Ministry of Agriculture of Yangzhou University Stored and provided by the Open Lab.

[0047] PCDNA3.1(+) and Escherichia coli DH5α were preserved and provided by Jiangsu Key Laboratory of Animal Preventive Medicine, Yangzhou University. Colloidal gold test strip preparation materials were purchased from Shanghai Gold Standard Biotechnology Co., Ltd.

[0048] The specific operation steps are as follows:

[0049] 1. Isolation and purification of CCV virus

[0050] (1) Isolation of CCV virus

[0051] Take fresh CCV disease ...

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Abstract

The invention relates to a CCV (canine corona virus) M protein monoclonal antibody and a preparation method thereof, and a preparation method of an immune colloidal gold test strip and particularly relates to a CCV M protein monoclonal antibody, comprising CCV M protein specific monoclonal antibodies 2H11 and 2G3. The preparation method of the CCV M protein monoclonal antibody comprises: preparinga CCV monoclonal antibody with separated and purified inactivated CCV as an immunogen by means of a monoclonal antibody technique; verifying the monoclonal antibody for the M protein by means of western-blot test, performing genetic cloning on the CCV M protein by means of recombinase ExnaseTMII to obtain a linear pCDNA3.1 vector, transfecting with 293T cells to carry out M protein expression, and carrying out IFA test verification to obtain two CCV M protein specific monoclonal antibodies 2H11 and 2G3. The invention also discloses the preparation method of the immune colloidal gold test strip. The CCV M protein monoclonal antibody herein has high specificity, high sensitivity, good operational simplicity and low cost, is applicable to clinical and field tests and is applicable to CCV etiological diagnosis, epidemiological investigation and quick diagnosis in animal hospitals.

Description

technical field [0001] The invention relates to a monoclonal antibody of CCV virus M protein and a preparation method thereof, and a preparation method of a canine coronavirus colloidal gold detection test strip, belonging to the fields of biotechnology and animal disease quarantine detection. Background technique [0002] Canine coronavirus (CCV) belongs to the genus Coronaviridae of the family Coronaviridae. The morphological characteristics of the virus include crown-like filaments, a capsule, a round or oval shape, and a size of 80-120nm. Canine coronavirus mainly has 4 kinds of structural proteins: spike protein (S, Spike Protein, 180-220kD) is the receptor binding site; small envelope glycoprotein (E, Envelope Protein, 9-12kD), this protein is relatively Small and envelope binding; transmembrane protein M protein (M, Transmembrane Protein, 29-32kD) is responsible for the transmembrane transport of nutrients and the budding release of newborn viruses and the formation o...

Claims

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Application Information

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IPC IPC(8): C07K16/10G01N33/569G01N33/558G01N33/52
CPCC07K16/10G01N33/56983G01N33/558G01N33/52
Inventor 甘军纪田晓彦吕海峰刘秀梵
Owner YANGZHOU UNIV
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