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Brucella antibody competitive AlphaLISA detection kit and detection method thereof

A detection kit, Brucella technology, applied in the fields of veterinary medicine, immunology and biological products, to ensure the effect of authenticity

Inactive Publication Date: 2019-01-01
CHONGQING UNIV OF TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are few reports at home and abroad on the detection of Brucella AlphaLISA

Method used

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  • Brucella antibody competitive AlphaLISA detection kit and detection method thereof
  • Brucella antibody competitive AlphaLISA detection kit and detection method thereof
  • Brucella antibody competitive AlphaLISA detection kit and detection method thereof

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Experimental program
Comparison scheme
Effect test

Embodiment

[0042] 1. Antigen screening

[0043] Studies have shown that after knocking out the Omp16 gene, Brucella affected the connection of the outer membrane peptidoglycan, resulting in damage to the structure of the outer membrane of Brucella, and finally increased the permeability of the outer membrane, resulting in cell lysis and death, indicating that OMP16 It is an outer membrane protein essential for the survival of Brucella. In addition, OMP16 protein is a new molecular pattern associated with bacterial pathogens, which can activate intracellular dendritic cells and induce Th1-type immune responses. It is a promising self-adjuvant against systemic and acquired brucellosis. Vaccines, Brucella proteins that achieve the same level of immune protection as live attenuated vaccines without the need for external adjuvants. Moreover, the OMP16 protein can be expressed in all Brucella species and has wide applicability, so the OMP16 protein is selected as the antigen.

[0044] 2. Pro...

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Abstract

The invention discloses a Brucella antibody competition AlphaLISA detection kit and a detection method thereof. The kit comprises a Brucella OMP16 protein monoclonal antibody, goat anti-mouse IgG-coupled receptor microbeads, His-tagged Brucella OMP16 protein antigen and nickel-chelated donor magnetic beads. The outer membrane protein of Brucella OMP16 is prepared by a genetic engineering technology, the Brucella OMP16 protein monoclonal antibody is prepared by a PEG1500 cell fusion technology, the purified recombinant OMP16 protein used as an antigen is interacted with the prepared monoclonalantibody, and the detection method of competitive Brucella antibody AlphaLISA is established by competitively binding the recombinant OMP16 protein to the antibody and the monoclonal antibody in a sample. The detection method has the advantages of short detection time, good specificity, high sensitivity, no need for washing, simple operation and low detection cost.

Description

technical field [0001] The invention relates to the fields of veterinary medicine, immunology and biological products, in particular to a Brucella antibody competition AlphaLISA detection kit and a detection method thereof. Background technique [0002] Brucellosis, referred to as brucellosis, is one of the most widespread and harmful zoonotic diseases in the world caused by Brucella. It is characterized by miscarriage, fever, and joint damage, threatening the life and health of humans and more than 60 species of mammals. "Animal Epidemic Prevention Law of the People's Republic of China", "National Medium and Long-Term Animal Disease Prevention and Control Plan (2012-2020)", "Chongqing Municipal Animal Epidemic Prevention Regulations", "Chongqing Medium and Long-term Animal Disease Prevention and Control Plan (2012-2020)" all stipulate In order to deal with brucellosis and carry out timely control. Therefore, early, rapid and accurate diagnosis of the disease is the key to...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/68
Inventor 吴胜昔曾政黄恒李俊萱黄诚李志徐缘李令臣鲁友铭侯力嘉曹芸董春霞
Owner CHONGQING UNIV OF TECH
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