Group 4 avian adenovirus sd-f strain, inactivated vaccine and preparation method

A poultry adenovirus, SD-F technology, applied in the field of bioengineering, can solve the problems that hinder the research process of poultry adenovirus vaccines, the tumorigenicity of cells, the risk of vaccine tumorigenesis, etc., and achieve no tumorigenic risk, simple operation, and low cost Effect

Active Publication Date: 2022-08-09
JIANGSU ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In large-scale production of chicken embryo proliferation, the process is cumbersome and expensive. Although tumorigenic LMH can be used to proliferate, LMH cells are tumorigenic, which poses a potential tumorigenic risk to vaccine production, which seriously hinders avian adenovirus vaccines. research process

Method used

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  • Group 4 avian adenovirus sd-f strain, inactivated vaccine and preparation method
  • Group 4 avian adenovirus sd-f strain, inactivated vaccine and preparation method
  • Group 4 avian adenovirus sd-f strain, inactivated vaccine and preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] This example describes the acquisition and specific identification of the SD-F strain of group I avian adenovirus type 4.

[0022] SPF eggs were purchased from Beijing Boehringer Ingelheim Viton Biotechnology Co., Ltd. and hatched to 10 days of age.

[0023] DF-1 cells were purchased from ATCC, USA.

[0024] MEM cell culture medium, DMEM medium, calf serum, trypsin were purchased from GIBCO Company.

[0025] Preparation of chicken embryo fibroblasts: CEFs were prepared according to the appendix of the 2000 edition of the Regulations for Veterinary Biological Products of the People's Republic of China.

[0026] 1. Obtainment of SD-F strain of group Ⅰ avian adenovirus type 4

[0027] Group Ⅰ avian adenovirus type 4 SD2015 strain, deposited in the China Center for Type Culture Collection, the deposit number is CCTCC NO: V201645.

[0028] After thawing the SD2015 strain of group I avian adenovirus type 4, centrifuge at 12,000 rpm for 10 minutes, and take the supernatant ...

Embodiment 2

[0065] Example 2 Preparation of avian adenovirus inactivated vaccine

[0066] The preparation method of avian adenovirus inactivated vaccine comprises the following steps:

[0067] (1) Culture of DF-1 cells

[0068] The formula of cell growth medium: add calf serum with a final concentration of 5% (volume percentage) and penicillin-streptomycin solution with a final concentration of 1% (volume percentage) to the MEM culture medium, and adjust the pH to After 7.0 to 7.2, use 0.22um filter paper to filter and pack and set aside. The penicillin-streptomycin solution is an aqueous solution containing 1000 U / L penicillin and 1000 U / L streptomycin.

[0069] The DF-1 seed cells were taken out from liquid nitrogen, thawed quickly in a 37°C water bath, and cultured in cell growth medium for 2 to 3 days for recovery. 7 ) were passaged 1:3.

[0070] (2) Virus inoculation: After the passaged DF-1 cells in step (1) grew into a dense monolayer, the cell surface was washed once with PBS,...

Embodiment 3

[0078] Example 3 Immune efficacy of inactivated vaccine against SD-F strain of avian adenovirus type 4 of group I

[0079] According to the method of Example 2, three batches of inactivated vaccines of group I type 4 avian adenovirus SD-F strain (20170210, 20170211, 20170212 batches of vaccines) were prepared. The virus content of each batch of vaccines is ≥ 10 4.4 / Feather portion.

[0080] For each batch of vaccine, 20 21-day-old SPF chickens were taken as the immunization group, and 0.3 ml of inactivated vaccine of group I group 4 avian adenovirus SD-F strain was subcutaneously injected into the neck of each chicken; the other 20 chickens were used as control and were not immunized. From 21 to 28 days after immunization, 10 animals in the immunized group and the control group were challenged, and the challenge dose was 10 7.0 TCID 50 (0.3ml) of liver tissue toxicity. Among them, the preparation method of liver tissue virus is as follows: Inject 49-day-old SPF chickens ...

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Abstract

The invention provides SD-F strain of group I type 4 avian adenovirus, an inactivated vaccine and a preparation method, and belongs to the field of bioengineering. Group I type 4 avian adenovirus SD-F strain, deposit number: CCTCC NO: V201964. The preparation method of the avian adenovirus inactivated vaccine is as follows: inactivate the SD-F strain virus liquid of group I 4 avian adenovirus, then mix with Tween-80, stir and mix to obtain an aqueous phase; This-80 is mixed, stirred evenly, and an oil phase is obtained after sterilization; the water phase and the oil phase are mixed in a volume ratio of 1:2-4, and emulsified to obtain an inactivated vaccine. The SD-F strain of the invention can adapt to non-tumorigenic DF-1 cells, has simple preparation process, low cost, no tumorigenic risk, and high safety; the inactivated vaccine has a high immune protection rate, and the antibody is positive 9 days after immunization The rate of infection was over 90%, the duration of immunization was 6 months, and the protection rate of progeny against virus was over 90%, and the protection rate against virus was 100%.

Description

technical field [0001] The invention relates to an avian adenovirus plaque cloned and purified strain, an inactivated vaccine and a preparation method, and belongs to the field of biological engineering. Background technique [0002] Avian adenovirus is a newly popular acute contact infectious disease. The infection is more common in chicken flocks, and even has a national epidemic, and it is increasing year by year. The disease is mainly transmitted vertically, but can also be transmitted horizontally. After infection, chickens can become life-long carriers and can be intermittently detoxified. The host range of the disease is getting wider and wider, and white-feathered broilers, laying hens, and yellow-feathered chickens can all be infected. The mortality rate of this disease is 20% to 80%, which brings huge economic losses to the breeding industry. Since this disease is a new disease, there are few commercial vaccines, and the most effective way to control this disease...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/00A61K39/235A61P31/20C12R1/93
CPCC12N7/00A61K39/12A61P31/20C12N2710/10221C12N2710/10234A61K2039/5252A61K2039/552Y02A50/30
Inventor 揭鸿英于漾毕志香赵阳朱亚露何家惠侯继波
Owner JIANGSU ACAD OF AGRI SCI
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