Inactivated vaccine for preventing and treating novel goose astroviruses and preparation method thereof
An astrovirus, inactivated vaccine technology, applied in the directions of biochemical equipment and methods, viruses, antiviral agents, etc., can solve the ineffectiveness of antiviral and antibacterial treatment methods, the reduction of the qualified rate of meat goose slaughter, and the economical economy of meat goose breeding industry. Loss and other problems, to achieve the effect of good commercial development prospects, good safety, and effective immune protection
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[0035] In the preparation method of the above-mentioned inactivated vaccine, GKC cells are used to cultivate the virus. This cell line has very good susceptibility to the novel goose astrovirus of the present invention, can be continuously passed down, and can also be cultured in suspension. Virus multiplication and vaccine production.
[0036] Adjuvants refer to substances that can effectively promote, enhance or prolong the body's specific immune response to antigens after being added to vaccine preparations. The selection of adjuvants is the basis of the vaccine emulsification process, and a stable vaccine emulsion can effectively improve the ability of animals to resist diseases. Therefore, choosing the appropriate adjuvant and determining the optimum ratio of water phase and oil phase are of great significance to the stability of the vaccine.
Embodiment 1
[0039] Example 1: Discovery and Identification of Novel Goose Astrovirus
[0040] 1. Materials and methods
[0041] 1.1 Collection and processing of disease materials:
[0042] Take liver, spleen, kidney and other tissues of dead goslings, add sterile saline at a ratio of 1:4 for homogenization, place the homogenate in the refrigerator for 3 times, centrifuge at 6000r / min for 15min, and take the supernatant The solution was filtered through a 0.22 μm filter and stored at -20°C for later use.
[0043] 1.2 Main reagents:
[0044] Goose embryos were purchased from a goose farm in Liaocheng, Shandong, the RNA extraction kit was purchased from Beijing Kangwei Century Co., Ltd., the ordinary agarose gel recovery kit was purchased from Beijing Quanshijin Company, and the reverse transcription kit was purchased from Bao Biology (Dalian ) Co., Ltd., 2×EsTaq MasterMix was purchased from Beijing Kangwei Century Co., Ltd., and DL2000Marker was purchased from Bao Biological (Dalian) Co....
Embodiment 2
[0066] Embodiment 2: the preparation of inactivated vaccine
[0067] (1) Virus propagation and harvest:
[0068] The isolated and identified novel goose astrovirus was inoculated into well-growing GKC cells, and the virus was proliferated in large quantities to obtain sufficient virus liquid. The obtained sufficient amount of cell virus liquid was frozen at -20°C, and after two freezing and thawing, the cell virus liquid was collected.
[0069] (2) Purification of virus:
[0070] Use established detection methods such as PCR and RT-PCR to detect whether other common viruses are contained in the obtained virus liquid. The detection items include avian influenza virus (AIV), Newcastle disease virus (NDV), goose parvovirus (GPV), infectious bronchitis virus (IBV), infectious bursal virus (IBDV), etc., and through the use of corresponding antibodies and The virus liquid is neutralized and the seed virus is purified.
[0071] (3) Inactivation of virus liquid:
[0072] The viru...
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