Preparation method of chicken infectious bursal disease virus and aviadenovirus IV combined inactivate vaccine

A dual inactivated vaccine and chicken infectious technology, which is applied in the field of veterinary biological products, can solve the problems of producing bursa and avian adenovirus type 4 dual inactivated vaccines, and achieve complete immune protection without external The source of pathogens and antibodies produces fast results

Inactive Publication Date: 2017-06-13
广州博恒生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] There is currently no report on the use of LMH cell lines to produce dual inactivated vaccines against bursa and avian adenovirus type 4

Method used

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  • Preparation method of chicken infectious bursal disease virus and aviadenovirus IV combined inactivate vaccine
  • Preparation method of chicken infectious bursal disease virus and aviadenovirus IV combined inactivate vaccine
  • Preparation method of chicken infectious bursal disease virus and aviadenovirus IV combined inactivate vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] The preparation of embodiment 1 liposome complex

[0040] (1) Weigh 0.2g of phosphatidylcholine, 0.08g of cholesterol, and 0.04g of distearoylphosphatidylethanolamine, mix them, dissolve in 50mL of ethanol, and sonicate for 5min;

[0041] (2) Rotate evaporation at 28°C and 0.1MPa to remove ethanol, let stand for 30min, add 100mL phosphate buffer solution containing 0.5% (m / v) catalase at pH 7.4, and place in a water bath at 35°C Medium shaking hydration reaction for about 1 hour to obtain liposome suspension;

[0042] (3) Adjust the particle size of the liposome suspension to 150 nm using a syringe filter to obtain liposomes containing catalase inside.

Embodiment 2

[0043] Example 2 Preparation of Chicken Infectious Bursal of Fabricius and Avian Adenovirus Type 4 Seedling Virus Liquid and Determination of Virus Content

[0044] (1) Preparation of seedling virus liquid:

[0045] ①Preparation of passage cell line LMH cells: LMH cells were digested and subcultured with 0.025% trypsin-0.02% EDTA with a mass volume ratio of 0.025%, added with 10% (v / v) newborn bovine serum, 1.0% (v / v ) double antibody and DMEM culture fluid of 2.0% (m / v) hydroxyethylpiperazineethanesulfonic acid, at 37 ℃, 5% CO 2 Cultivate in an incubator until a well-growing monolayer of cells is formed, and then wash the cells with serum-free DMEM medium for 3 times for inoculation of chicken infectious bursal virus and avian adenovirus type 4.

[0046] ② Seed virus propagation: inoculate chicken infectious bursal virus seeds into the cells prepared in step ① at a final volume of 1:100, and inoculate avian adenovirus type 4 virus seeds into the cells prepared in step ① at a...

Embodiment 3

[0050] Embodiment 3 Inactivation and inactivation test of chicken infectious bursa and avian adenovirus type 4 seedling virus liquid

[0051] (1) Inactivation of chicken infectious bursa / fowl adenovirus type 4: the virus liquid is imported in the inactivation tank, added formalin solution, fully mixed, the final concentration of formalin solution is 0.1%, After inactivation at 37°C for 16 hours (the timing starts when the temperature in the tank reaches 37°C), take it out and store it at 6°C, which should not exceed 1 month.

[0052] (2) chicken infectious bursal / fowl adenovirus type 4 virus inactivation test: get the sample of inactivation test, use DMEM nutrient solution with 10 -1 、10 -2 、10 -3 Dilute the samples to be tested in three proportions, and set up inactivated pre-virus samples with the same dilution ratio as positive controls, inoculate them in 48 wells of LMH cell monolayer cells, and inoculate 5 wells in each group, 0.4mL per well at 37°C, 5 %CO 2 Incubate ...

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Abstract

The invention relates to a method for producing chicken infectious bursal and avian adenovirus type 4 inactivated vaccines with LMH cell lines, which comprises the following steps: separately injecting chicken infectious bursal and avian adenovirus type 4 viruses Inoculate in LMH cells, obtain chicken infectious bursa seedling virus liquid and avian adenovirus type 4 seedling virus liquid by culturing, freezing and thawing, centrifuging, concentrating and other steps, and then sterilize the seedling virus liquid of the two viruses After inactivation, equal volumes are mixed and emulsified to make an emulsion-type inactivated vaccine. The dual vaccine provided by the invention has high potency, has complete immune protection against chicken infectious bursal virus and avian adenovirus type 4, has good safety, is stable and effective, and has a long protection period.

Description

technical field [0001] The invention belongs to the technical field of veterinary biological products, and in particular relates to a method for producing chicken infectious bursa and avian adenovirus type 4 dual inactivated vaccines by using LMH cell lines. Background technique [0002] Chicken infectious bursal disease is an acute and contagious disease of chickens caused by Reovirus. Due to the sudden onset of the disease, short course of disease, high mortality, and the immunosuppression of chickens, it is still one of the main infectious diseases in the chicken industry. There are two types of vaccines for bursal virus on the market: live vaccines and inactivated vaccines. Live vaccines with weak virulence have poor immune effect, and those with strong virulence can damage the bursa of Fabricius and affect the immune effect; inactivated vaccines retain the complete virus structure and greatly improve the safety performance without affecting the immune effect. [0003]...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/12A61K39/235A61K9/107A61P31/14A61P31/20C12N7/00C12R1/93
CPCA61K39/12A61K9/107A61K2039/5252A61K2039/552A61K2039/70C12N7/00C12N2710/10234C12N2710/10251C12N2720/10034C12N2720/10051
Inventor 张毓金谢秉超严悌昆
Owner 广州博恒生物科技有限公司
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