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Monoclonal antibody, testing kit including the monoclonal antibody and application

A technology of monoclonal antibody and kit, applied in the field of immune application, achieving the effect of low temperature requirement, short detection time and easy operation

Inactive Publication Date: 2005-10-26
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In terms of diagnosis and detection of chicken epidemics, Chinese Invention Patent Publication No. 99101537.1 discloses the preparation and application of a quick diagnostic test strip for livestock and poultry diseases, but the main point of this invention is the detection of pathogens of livestock and poultry diseases, and does not involve the detection of Detection of Antibody of Chicken Avian Influenza Virus

Method used

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  • Monoclonal antibody, testing kit including the monoclonal antibody and application
  • Monoclonal antibody, testing kit including the monoclonal antibody and application
  • Monoclonal antibody, testing kit including the monoclonal antibody and application

Examples

Experimental program
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Effect test

Embodiment 1

[0026] 1. Preparation of anti-chicken IgG Fc monoclonal antibody:

[0027] With reference to Xue Qingshan's "Principles and Techniques of In Vitro Culture" Science Press, 2001 edition, the method in: Utilize the chicken IgG extracted by the inventor's Microbiology and Immunology Laboratory to immunize Balb / C mice (purchased from the Experimental Animal Center of Hubei Academy of Medical Sciences) ), the immunization procedure was to emulsify a protein solution containing 50-100 μg of chicken IgG with an equal volume of Freund's complete adjuvant (purchased from sigma company), inject it into the abdominal cavity of mice, and then strengthen it every ten days, and replace it with incomplete adjuvant Agent emulsification (sigma). Finally, three days before the fusion, (preferably more than 4 weeks after the last immunization), intraperitoneal booster immunization, double the amount of antigen, without adjuvant. At the time of fusion, take a Balb / C mouse that has undergone the f...

Embodiment 2

[0031] Preparation of Chicken Avian Influenza Virus NP Protein:

[0032] For the molecular biology methods used, please refer to the literature: Sambrook J, Fritsch EF, Mani Artis T, editors (translated by Jin Dongyan, Li Mengfeng, etc.). A Laboratory Guide to Molecular Cloning. The second edition, Beijing Science Press, 1992 provided the method.

[0033] 1. Cloning and sequencing of NP gene

[0034] The NP gene involved in the present invention was cloned by the applicant himself, and the sequence of the gene has been registered in the GeneBank database with the accession number AY788915. The sequence has 97% homology with known sequences. The specific method of cloning and sequencing of NP gene is: a strain A / chicken / China / HSS2004 (H9N2) [GeneBank accession number is AY788915, the isolation and For identification, see literature: Li Zili et al. Diagnosis of avian influenza in Hubei Province and isolation and preliminary identification of virus strains, Chinese Journal of...

Embodiment 3

[0040] Preparation of rabbit anti-mouse IgG antibody:

[0041] Refer to editor-in-chief such as He Zhaoyang, "Animal Immunology Experimental Technology" Changchun: Jilin Science and Technology Press, the method in 2002: Utilize the Balb / C mouse IgG that the inventor's place extracts from the microbiology and immunity laboratory to immunize healthy rabbits, prepare high Specific and high-titer rabbit anti-mouse IgG hyperimmune serum, the hyperimmune serum is crudely extracted by saturated ammonium sulfate precipitation, and high-purity rabbit anti-mouse IgG antibody is obtained after passing through the G-200 column. Use this antibody as a quality control line.

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Abstract

The present invention discloses a reagent box for quickly detect chicken and birds flu virus antibody. This reagent box includes body, 96 aperture enzyme mark bar in the box, test paper and sample diluted liquid. The test paper consist of sample cushion, absorb cushion, besmear IgG Fc golden cushion, bundled with NP decect string and IgG quality string pyroxylin film, in turn absorb cushion, pyroxylin film, golden cushion, sample cushion plaster unsopped water supported slice. This invention relates to a kind of special monoclonal antibody cross bred tumour cell BDRPDP and special identify IgGFc monoclonal antibody preparation. The reagent box which detect chicken and birds flu virus has strong diathesis, higher sensitivity, simply manipulation and diagnose quickly distinct characteristic.

Description

technical field [0001] The invention belongs to the field of immune application and relates to related fields such as animal molecular biology and immunology. Specifically, the present invention relates to the preparation of a specific monoclonal antibody, a kit containing the monoclonal antibody and its application in the diagnosis and detection of chicken avian influenza virus antibody. Background technique [0002] As we all know, avian influenza, Newcastle disease, bursa, infectious bronchitis, infectious laryngotracheitis, Marek, egg drop syndrome and other diseases are important diseases that endanger the global poultry industry, especially avian influenza. At the beginning of 2004, H5N1 subtype highly pathogenic avian influenza occurred one after another in some Asian countries and regions. The poultry industry in the region has caused huge economic losses, so the governments of various Asian countries have attached great importance to the prevention and control of b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/16
Inventor 毕丁仁彭大鹏李自力华艳胡思顺肖运才石德时刘梅许清荣
Owner HUAZHONG AGRI UNIV
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