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Detection method of aGST-ELISA of avian influenza virus (H5N1 subset) serum antibody

A technology of avian influenza virus and detection method, which is applied in the field of aGST-ELISA detection of avian influenza virus serum antibodies, can solve the problems of reduced sensitivity of detection methods, and that the product purity cannot meet the requirements of establishing ELISA.

Inactive Publication Date: 2006-11-29
北京翎羽生物科技有限公司
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Problems solved by technology

However, since the prokaryotic expression product of the HA gene is an insoluble protein, denaturation and renaturation are required to obtain the purified product, and the spatial structure of the expression product often changes to a certain extent after denaturation and renaturation, resulting in the sensitivity of the detection method. In addition, the purity of the product obtained by this purification method cannot meet the requirements for establishing an ELISA

Method used

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  • Detection method of aGST-ELISA of avian influenza virus (H5N1 subset) serum antibody
  • Detection method of aGST-ELISA of avian influenza virus (H5N1 subset) serum antibody
  • Detection method of aGST-ELISA of avian influenza virus (H5N1 subset) serum antibody

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Embodiment

[0030] 1. Experimental materials

[0031] 1. Hybridoma cell line secreting anti-glutathione-sulfur-transferase (GST) monoclonal antibody: developed and preserved by the High-Tech Laboratory of the Institute of Animal Husbandry and Veterinary Medicine, Beijing Academy of Agriculture and Forestry, and available for sale. After several passages, cryopreservation and recovery of the cell line, the hybridoma cells secreted antibodies stably, the cells grew vigorously, and the morphology was good. cell growth state. (like figure 2 ) The anti-GST monoclonal antibody used in the present invention is prepared by itself, and this product can be purchased from the market.

[0032] 2. AIV HA1 genetically engineered recombinant expression bacteria: the HA1 cloned gene of the hemagglutinin main antigen part of avian influenza virus A / goose / Guangdong / 96 (H5N1) was cloned into the expression plasmid pGEX-5x-1 to construct a recombinant plasmid (Zhou Xuemei, Huo Huiling , Zhang Biao, Li Yo...

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Abstract

The invention relates to 'an aGST-ELISA detection method for bird flu viruses (H5N1 sub) serum antibodies', and its detection antigen is high- purity fusion protein GST-HA1 made by capturing fusion protein GST-HA1 on enzyme link plate by anti-GST monoantibody. And the method can be used to detect bird flu serum antibodies. And it has characters of simple antigen preparing process, high biosafety, low cost, easy to produce, etc, and lays the foundation of further development and research of bird flu detection reagent box.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to aGST-ELISA detection method of avian influenza virus serum antibodies. Background technique [0002] Avian Influenza (AI) is an infectious disease or infection syndrome of birds, which was first reported by Perroncito in Italy in 1878, and is called chicken plague. In 1955 Shafer confirmed that the disease was caused by influenza A virus. For more than 100 years, the epidemics or outbreaks of avian influenza caused by specific strains around the world have caused harm to the poultry industry and human health, especially the highly pathogenic avian influenza, because of its rapid spread and high mortality, Due to its great harm to poultry production and its important public health significance, it has been widely valued by the world. Therefore, it is classified as a Class A infectious disease by the International Bureau of Animal Epidemiology (OIE), and my country cla...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/543
Inventor 李永清张飚黄金海王宏俊
Owner 北京翎羽生物科技有限公司
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