Recombined pig influenza virus NP antigen, its preparing method and use in diagnosis

A technology for swine influenza virus and influenza virus infection, which is applied in the field of diagnostic antigens to achieve the effects of good specificity and sensitivity, easy promotion and use, and easy production.

Inactive Publication Date: 2003-11-26
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Thereafter recovers and develops a strong and long-lasting immunity, but also becomes a long-term carrier and detoxifier

Method used

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  • Recombined pig influenza virus NP antigen, its preparing method and use in diagnosis
  • Recombined pig influenza virus NP antigen, its preparing method and use in diagnosis
  • Recombined pig influenza virus NP antigen, its preparing method and use in diagnosis

Examples

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Embodiment 1

[0026] Recombinant swine influenza virus NP antigen, the antigen is a recombinant nucleoprotein antigen for diagnosing type A influenza virus infection, it adopts Escherichia coli prokaryotic expression system to express the complete NP of my country's swine influenza virus (SIV) isolate (H3N2 subtype) Gene, using the expressed NP protein as an antigen to detect the anti-NP protein antibody produced by influenza A virus infection.

Embodiment 2

[0028] The recombinant swine influenza virus NP antigen expresses the NP protein of swine influenza virus H3N2 subtype by using an Escherichia coli expression system, and is used as an antigen for diagnosis of type A influenza virus infection after purification.

Embodiment 3

[0030] A method for cultivating recombinant swine influenza virus NP antigen, using RT-PCR method to clone the NP gene isolated from H3N2 subtype swine influenza virus, then digesting the complete NP gene with restriction endonuclease and then combining it with the same treatment The prokaryotic expression vector pET30 was connected to obtain the expression plasmid pETNP containing the SIV NP gene;

[0031] The recipient strain BL21 was transformed with pETNP, a single colony was picked, induced and acclimatized with different concentrations of inducers, and samples were collected at different times. The expression of NP was confirmed by SDS-PAGE electrophoresis analysis, and the optimal induction concentration and induction time of IPTG were determined to be 1mM and 6 hours respectively;

[0032] The protein band after SDS-PAGE was transferred to the membrane, and the Western-blot test confirmed that the expressed recombinant nucleoprotein was of the same size and had reactiv...

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Abstract

The present invention discloses recombinant pig influenza virus NP antigen and its preparation process and application in diagnosis. Type-A influenza virus genome consists of 8 segments of RNA, and its nucleoprotein (NP) has type and group specificity and is the basis of typing diagnosis. The recombinant pig influenza virus NP antigen has colibacillus prokaryotic expression system to express complete NP gene of separated pig influenza virus strain in China, and the expressed NP is used as antigen in detecting type-A influenza virus infection produced anti-NP protein antibody. The NP gene is the basis for classifying and diagnosing pig influenza virus with many subtypes. Therefore, the diagnosis technology established based on recombinant SIV NP antigen may be used in detecting type-A influenza virus infection of pig, fowl, horse and human.

Description

[0001] Technical field: the present invention relates to the field of veterinary biotechnology, using the Escherichia coli expression system to express the NP protein of swine influenza virus (SIV) H3N2, and use it as an antigen for diagnosis after purification. Background technique: [0002] Swine Influenza (Swine Influenza) is an acute respiratory infectious disease of pigs caused by swine influenza A virus of the family Orthomyxoviridae, characterized by sudden onset, rapid spread, shortness of breath and paroxysmal cough. After that, they can recover and develop strong and lasting immunity, but they also become long-term carriers and detoxifiers. Swine flu was first reported in the United States in 1918 and is now believed to be related to the 1918 human flu pandemic. In 1930, Shope and others isolated the first strain of H1N1 subtype swine influenza from Iowa. After that, various subtypes of influenza viruses were isolated from pigs. Swine influenza is endemic and globa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/11C12N15/44C12N15/70G01N33/531G01N33/569
Inventor 童光志倪健强李海燕仇华吉王云峰
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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