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Recombination H1N1 subtype swine flu bivalent inactivated vaccine and preparation method and application thereof

A bivalent inactivated vaccine, H1N1 technology, applied in the field of recombinant H1N1 subtype swine influenza bivalent inactivated vaccine and its preparation, can solve the problems of difficult handling of chicken embryos, high consumption, antigenic variation, etc., and achieve benefits to the industry The effect of globalization and market application

Inactive Publication Date: 2017-11-24
SHANGHAI VETERINARY RES INST CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although chicken embryo inoculation to produce vaccines is easy to operate and has rich storage experience, it inevitably brings disadvantages such as large time-consuming, high consumption, easy pollution in the production process, difficult quality assurance, and difficult handling of infected chicken embryos. Influenza virus often undergoes antigenic variation during inoculation in chicken embryos, which affects the vaccine effect
If the circulating virus is a highly pathogenic avian influenza virus, inoculation of chicken embryos will lead to a large number of deaths and seriously affect the production of normal vaccines (Takada A et al., 1999)

Method used

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  • Recombination H1N1 subtype swine flu bivalent inactivated vaccine and preparation method and application thereof
  • Recombination H1N1 subtype swine flu bivalent inactivated vaccine and preparation method and application thereof
  • Recombination H1N1 subtype swine flu bivalent inactivated vaccine and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 Construction of European Avian H1N1 Subtype SIV Recombinant Strain SH1 / PR8 and Classical H1N1 Subtype SIV Recombinant Strain G11 / PR8

[0030] Using European poultry H1N1 subtype swine influenza virus domestic epidemic strain A / swine / Shanghai / 1 / 2014 (H1N1) (SH1) as material, using RT-PCR to amplify HA and NA genes (SEQ ID NO.3 and SEQ ID NO.3 and NO.4), and clone it into a PBD vector to construct recombinant plasmids of HA and NA genes. The HA and NA gene recombinant plasmids and 6 internal gene (PB2, PB1, PA, NP, M and NS) recombinant plasmids derived from influenza virus strain A / Puerto Rico / 8 / 1934 (H1N1) (PR8) were co-transformed Infected 293T cells, and successfully constructed the recombinant European avian H1N1 subtype swine influenza vaccine strain SH1 / PR8. The recombinant vaccine strain SH1 / PR8 has higher virus titer and hemagglutination titer on MDCK cells or chicken embryos.

[0031] Using the classical H1N1 subtype swine influenza virus domestic ep...

Embodiment 2

[0032] Example 2 Preparation of Recombinant H1N1 Subtype Swine Influenza Bivalent Inactivated Vaccine

[0033] The recombinant European avian H1N1 subtype swine influenza virus (SH / PR8) and the classical H1N1 subtype swine influenza virus (G11 / PR8) were proliferated in large quantities on MDCK cells. The proliferated virus was inactivated by 0.1% formaldehyde solution at 37°C for 48 hours, and the HA hemagglutination titer was detected after the blind passage of the inactivated virus for 3 generations (if the HA hemagglutination titer cannot be detected, it means that the inactivated virus is safe, can enter the next emulsification operation). Recombinant inactivated viruses SH / PR8 and G11 / PR8 were passed through oil-emulsion adjuvant (MONTANIDE TM ISA 61VG, Sepic Company, France) was emulsified, and the inactivated virus was emulsified with the weight ratio of adjuvant to antigen at 60:40 according to the requirements of the adjuvant instructions, and prepared into an inact...

Embodiment 3

[0034] Example 3 Evaluation of Immunoprotective Efficacy of Recombinant H1N1 Subtype Swine Influenza Bivalent Inactivated Vaccine

[0035] 1. Procedures for evaluation of bivalent inactivated vaccine immunization and challenge using mice as a model

[0036] (1) Choose healthy SPF female BALB / c mice at the age of 6-8 weeks, and divide them into 5 groups at random, with 11 each in Group 1, Group 2, Group 3, and Group 4, and 22 mice in Group 5 (11 of which are used for attacking). Toxicity test, 11 were only used as environmental controls). Raise and manage the mice according to the requirements of the SPF level, and raise the mice normally for 3 days to familiarize the mice with the feeding environment, and collect blood from the orbital vein to collect the mouse serum as a blank control;

[0037] (2) The specific immunization procedures of the mice in the experimental groups (Group 1, Group 2, Group 3, Group 4, and Group 5) are shown in Table 1 below. Immunize 200 μl of inact...

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Abstract

The invention discloses a recombination H1N1 subtype swine flu bivalent inactivated vaccine and further discloses a preparation method and application of the recombination H1N1 subtype swine flu bivalent inactivated vaccine. The recombination H1N1 subtype swine flu bivalent inactivated vaccine comprises a European avian origin H1N1 subtype swine flu virus and a classical H1N1 subtype swine flu virus antigen. According to the recombination H1N1 subtype swine flu bivalent inactivated vaccine, the high valence of a serum hemagglutination inhibition (HI) antibody, a neutralizing antibody and a specific (IgG) antibody can be generated by inducing the body, and 100% immune protection can be provided for attacking of European avian origin and classical H1N1 subtype swine flu viruses.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a recombinant H1N1 subtype swine influenza bivalent inactivated vaccine and its preparation method and application. Background technique [0002] Swine influenza is referred to as swine flu (Swine Influenza, SI), which is infected by influenza A virus of Orthomyxoviridae family, causing high fever, cough, dyspnea, and highly contagious respiratory infectious diseases in pigs. High, fast transmission, low mortality and other characteristics, not only seriously endanger the development of my country's pig industry, but also endanger human health (Shinde, Bridges et al., 2009), which has aroused the concern of global public health. Since human influenza virus receptors and avian influenza virus receptors exist on the surface of porcine respiratory epithelial cells, it is considered to be an important intermediate host for genetic reassortment and cross-species transmission of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/145A61P31/16
CPCA61K39/12A61K2039/5252A61K2039/552A61K2039/55566A61K2039/70C12N2760/16134A61K2300/00
Inventor 于海童光志阮宝阳宫晓倩刘晓敏李国新童武
Owner SHANGHAI VETERINARY RES INST CHINESE ACAD OF AGRI SCI
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