Inactivated vaccine for preventing and controlling new chicken reovirus and preparation method thereof
A technology of reovirus and inactivated vaccines, which is applied in the field of inactivated vaccines and preparations for the prevention and treatment of new chicken reoviruses, can solve the problems of ineffective antiviral and antibacterial treatment methods, and can not be prevented and controlled, so as to prevent and control infection and outbreak, simplify the production process, and realize the effect of large-scale production
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[0036] In the preparation method of the above-mentioned inactivated vaccine, the LMH cell line is used to culture the virus. The cell line has a very good susceptibility to the novel chicken reovirus of the present invention. It can be continuously passaged, and can also be cultured in suspension with good growth. Conducive to virus proliferation and vaccine production.
[0037] 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 choice of adjuvant is the basis of the vaccine emulsification process. A stable vaccine emulsion can effectively improve the ability of animals to resist diseases. Therefore, choosing a suitable adjuvant and determining the best ratio of water to oil are of great significance to the stability of the vaccine.
Embodiment 1
[0040] Example 1: Discovery and identification of new chicken reovirus
[0041] 1. Virus isolation:
[0042] (1) For arthritic chicks that were clinically identified as positive by PCR reaction, the swollen tendon tissue was collected by necropsy and placed in a 15mL centrifuge tube, and 5 times the volume of serum-free DMEM medium was added, homogenized and repeatedly frozen and thawed. 3 times, after each thawing, shake on the shaker for 1-2 minutes, and then perform the next freeze-thaw; after freezing and thawing, centrifuge the 15mL centrifuge tube containing the sample in the centrifuge at 4000rpm for 5min; take the supernatant, 0.22μm micropores After the filter membrane is filtered, spare;
[0043] (2) According to the proliferation characteristics of reo, chicken liver cancer cells (LMH) were selected for virus isolation. According to the conventional cell culture method, wait for 25cm 2 When the cells in the cell flask are covered with a monolayer, aspirate the culture me...
Embodiment 2
[0059] Example 2: Preparation of inactivated vaccine
[0060] (1) The propagation and harvest of viruses:
[0061] The isolated and identified new chicken reovirus is inoculated into a well-growing LMH cell line, and the virus is proliferated in large quantities to obtain sufficient virus liquid. The obtained sufficient cell virus liquid was frozen at -20°C, and the cell virus liquid was collected after two freeze-thaw cycles.
[0062] (2) Purification of virus:
[0063] Use established PCR, RT-PCR and other detection methods to detect whether the obtained virus liquid contains other common viruses. Test items include avian influenza virus (AIV), Newcastle disease virus (NDV), fowlpox virus (APV), Mycoplasma synovium (MS), infectious bronchitis virus (IBV) and infectious bursal disease virus (IBDV) Etc.; Centrifuge the harvested virus solution at 5000r / min for 5min to remove cell debris and other impurities, then use ultracentrifugation at 60000r / min for 2h, and resuspend with DMEM ...
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