Bovine viral diarrhoea virus inactivated vaccine and preparation method thereof
A technology for bovine viral diarrhea and inactivated vaccine, which is applied in the field of veterinary biological products and can solve problems such as economic losses
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Embodiment 1
[0063] ——Preparation of inactivated vaccine against bovine viral diarrhea
[0064] 1. Antigen Preparation
[0065] This embodiment provides the preparation of BVDV-L strain, BVDV-L2 strain, BVDV-T strain, and BVDV-D strain virus by using spinner bottle adherent culture, suspension culture technology, and microcarrier suspension culture technology to cultivate bovine kidney cell line (MDBK) Methods.
[0066] (1) Prepare BVDV-L strain, BVDV-L2 strain, BVDV-T strain, BVDV-D strain antigen by using spinner bottle adherent culture cells
[0067] 1) Cultivate cells: First, resuscitate the bovine kidney cell line MDBK into a cell flask, grow for about 48 hours, digest with EDTA-trypsin cell dispersion, and passage according to the volume ratio of 1:3. Add cell growth medium (DMEM medium containing 10% (v / v) calf serum, pH value 7.0) for spinner bottle culture, when the cells cover 90% to 100% of the monolayer, it is used to inoculate BVDV-L respectively strain, BVDV-L2 strain, BVD...
Embodiment 2
[0093] ——Inspection of finished products of bovine viral diarrhea inactivated vaccine
[0094] 1. Sterility test: according to the appendix of the current "Chinese Veterinary Pharmacopoeia", it should meet the requirements. The above test results are shown in Table 2.
[0095] Table 2 Physical properties, sterility test results
[0096]
[0097] 2. Safety inspection
[0098] (1) Alternative to animal testing
[0099] 1) Monovalent vaccine (BVDV I a inactivated vaccine, BVDV I b inactivated vaccine, BVDV II a inactivated vaccine and BVDV II b inactivated vaccine):
[0100] Each vaccine used 6 healthy rabbits of 1.5-2.0 kg, including 4 in the immunization group and 2 in the control group. The rabbits in the immunization group were injected with 1.0ml of vaccine in each leg muscle, and the rabbits in the control group were injected into each leg muscle. 1.0ml of cell culture was observed continuously for 7 days and body temperature was measured at fixed points every aftern...
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