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Preparation method of avian influenza virus and fowl adenovirus combined inactivated vaccine

A dual-inactivated vaccine and avian influenza virus technology, applied in biochemical equipment and methods, vaccines, viruses, etc., can solve the problems of immutable downstream technologies, low virus titers, and easy contamination, and achieve increased virus content, Improve the effect of vaccines and increase the effect of proliferation

Active Publication Date: 2017-08-18
广东渔跃生物技术有限公司 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, using SPF chicken embryo vectors for virus propagation, the virus titer is not high, and the production cost of large-scale production with SPF chicken embryo vectors is very expensive, which is not conducive to large-scale production
[0006] At the same time, inoculation of influenza virus through the allantoic cavity of chicken embryos will occur. For example, the supply of vaccines during a pandemic is often limited by the number of SPF chicken embryos; some strains cannot replicate in chicken embryos, so it is difficult to obtain a sufficient amount of virus, only chicken Embryo-adapted strains can obtain high-yield viruses; mutations in receptor-binding specific sites lead to reduced antigenicity; downstream technologies for vaccine production are immutable, heavy, and easy to contaminate, etc. Promotion and Application of Viral Vaccines

Method used

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  • Preparation method of avian influenza virus and fowl adenovirus combined inactivated vaccine
  • Preparation method of avian influenza virus and fowl adenovirus combined inactivated vaccine
  • Preparation method of avian influenza virus and fowl adenovirus combined inactivated vaccine

Examples

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Effect test

Embodiment 1

[0043] Embodiment 1, a kind of preparation method of avian influenza virus, avian adenovirus dual inactivated vaccine

[0044] Preparation of S1 cell carrier: Digest and disperse LMH cells with trypsin, and use DMEM medium containing 5% newborn bovine serum and 1000 units / mL penicillin-streptomycin double antibody at 37°C, 5% CO 2 LMH cells were cultured for 2 days under certain conditions, and then the cells were washed twice with serum-free DMEM medium to obtain cell carriers; the LMH cells were cultured in spinner bottles or placed in microcarrier reactors.

[0045] Inoculation of S2 virus: inoculate the virus liquid of avian influenza virus subtype H9 and avian adenovirus type 4 into the cell carrier obtained in step S1 at a final volume of 1:50, place it at 37°C for 30 minutes, and then suck out the virus liquid , followed by medium A, at 37°C, 5% CO 2 48h under the condition of culturing, 80% cytopathy occurs, and diseased cells are obtained;

[0046] Described medium ...

Embodiment 2

[0054] Embodiment 2, a kind of preparation method of avian influenza virus, avian adenovirus dual inactivated vaccine

[0055] Preparation of S1 cell carrier: Digest and disperse LMH cells with trypsin, and use DMEM medium containing 8% newborn bovine serum and 1500 units / mL penicillin-streptomycin double antibody at 37°C, 5% CO 2 The conditions were cultivated for 2 days, and then the cells were washed 3 times with serum-free DMEM culture medium to obtain cell carriers; the LMH cells were cultured in a microcarrier reactor, and the amount of microcarriers used was 6 g / L.

[0056] Inoculation of S2 virus: inoculate the virus liquid of avian influenza virus subtype H9 and avian adenovirus type 4 into the cell carrier obtained in step S1 at a final volume of 1:100, place it at 37°C for 40 minutes, and then suck out the virus liquid , followed by medium A, at 37°C, 5% CO 2 Under the conditions of 56 hours, 80% of the cells were damaged, and the diseased cells were obtained;

[...

Embodiment 3

[0065] Embodiment 3, a kind of preparation method of avian influenza virus, avian adenovirus dual inactivated vaccine

[0066] Preparation of S1 cell carrier: Digest and disperse LMH cells with trypsin, and incubate at 37°C, 5% CO with DMEM medium containing 10% newborn bovine serum and 2000 units / mL penicillin-streptomycin double antibody 2 The conditions were cultivated for 3 days, and then the cells were washed 3 times with serum-free DMEM culture medium to obtain cell carriers; the LMH cells were cultured in a microcarrier reactor, and the amount of microcarriers used was 8 grams per liter.

[0067] Inoculation of S2 virus: inoculate the virus liquid of avian influenza virus subtype H9 and avian adenovirus type 4 into the cell carrier obtained in step S1 at a final volume of 1:200, place it at 37°C for 60 minutes, and then suck out the virus liquid , followed by medium A, at 37°C, 5% CO 2 72h under the condition of culture, 80% cytopathy occurs, and diseased cells are obt...

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Abstract

The invention belongs to the technical field of veterinary biological products and particularly relates to a preparation method of avian influenza virus subtype H9 and fowl adenovirus 4 combined inactivated vaccine. LHM (Leghorn male hepatocarcinoma) continuous cell line is used as carrier cells to perform viral multiplication, culture medium A composed of glutamine, recombinant human insulin, human serum albumin, transferrin, biotin and growth factors is used to perform culturing, and a viral liquid is collected and inactivated to prepare the vaccine. The multiplication of avian influenza virus subtype H9 and fowl adenovirus 4 by the LHM continuous cell line has no need for additional pancreatin, the background of LHM cells is clear, no extraneous pathogens occur, the multiplication is easy, the process can be effectively simplified, and the cost can be reduced; in addition, the avian influenza virus and fowl adenovirus combined inactivated vaccine has high viral content, good stability and high safety and is an ideal avian influenza virus and fowl adenovirus combined inactivated vaccine.

Description

technical field [0001] The invention belongs to the technical field of veterinary biological products, and in particular relates to a preparation method of a dual inactivated vaccine of avian influenza virus H9 subtype and avian adenovirus type 4. Background technique [0002] H9N2 subtype avian influenza belongs to low pathogenicity avian influenza, but its incidence rate is high. Its main clinical manifestations are mild respiratory symptoms, reduced feed intake, egg production rate can drop from 90% to below 20%, or even stop production. About 30% of commercial broilers can die, and they are easily mixed with Escherichia coli, which seriously affects the production performance of poultry and brings serious economic losses to the poultry industry. Moreover, the H9N2 subtype AIV can pass through host barriers and infect mammals, including humans, and has the opportunity to spread in humans on a large scale, posing a serious threat to human health. [0003] Avian group I ad...

Claims

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Application Information

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IPC IPC(8): A61K39/295A61K39/235A61K39/145A61P31/16A61P31/20
CPCA61K39/12A61K2039/5252A61K2039/552A61K2039/70C12N2710/10234C12N2760/16134
Inventor 张毓金严悌昆谢秉超敖艳华
Owner 广东渔跃生物技术有限公司
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