Canine distemper parvovirus bigeminy subunit vaccine
A technology of subunit vaccines and parvoviruses, applied in the direction of vaccines, viruses, antiviral agents, etc., can solve the problems of short duration of immunity and high cost of inactivated vaccines, achieve good protection effects, prevent epidemics and spread, and reduce economic costs. loss effect
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Embodiment 1
[0019] Embodiment 1: Cloning expression of canine distemper virus H gene
[0020] The strains, bacterial strains, cells and plasmids required in this embodiment are as follows:
[0021] Both Vero cells and pFastBac1 plasmid were preserved by our laboratory; Escherichia coli E.coli DH5a was purchased from Treasure Bioengineering (Dalian) Co., Ltd.; Sf9 insect cells were donated by China Center for Animal Health and Epidemiology.
[0022] The H gene used in the present invention is amplified from the CDV QN-1 strain, and the QN-1 strain is a mink in a farm in Zhucheng City, Shandong Province, about 60 days old; unimmunized mink canine distemper vaccine product, mink The group had symptoms of suspected canine distemper, with obvious secretions from the eyes and nose, loss of appetite, and dry nose, but no mink deaths occurred, and it was suspected to be infected by the attenuated strain of mink distemper virus. Virus screening was performed on the samples obtained from the farm,...
Embodiment 3
[0055] Example 3: Laboratory trial production and product quality research of canine distemper virus H gene and canine parvo VP2 subunit vaccine
[0056] virus and cell
[0057] The virus seed used to manufacture this product is the recombinant baculovirus of CDV and CPV, which is constructed, kept and supplied by Qingdao Agricultural University, and the virus titer is ≥10 7 IFU / ml; the 6th passage of Sf9 production cells, identified, kept and supplied by Qingdao Agricultural University.
[0058] 1. Virus inoculation and cultivation
[0059] Inoculate the virus seeds at a 0.01% inoculum to a cell density of 1×10 6 Cells / ml Sf9 cells were continuously cultured at 27°C for 72 hours, the virus supernatant was harvested, centrifuged at 12,000rpm for 20 minutes, and the supernatant was collected, which was the virus for production, and stored at -20°C for later use.
[0060] When the Sf9 cell density in the bioreactor reaches 1×10 6 When cells / ml, the virus seeds for production...
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