Porcine pseudorabies virus, and vaccine composition and applications thereof
A technology for porcine pseudorabies virus and vaccine composition, which is applied in the directions of antiviral agents, viruses/phages, microorganisms, etc., can solve the problems of immune effect and unsatisfactory duration of antibodies, and achieves good immunogenicity, long duration, and antibodies. high level effect
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Embodiment 1
[0033] Example 1 The acquisition and characteristic determination of porcine pseudorabies virus
[0034] 1. Virus Isolation
[0035] The spleen, lung, kidney, and lymphoid tissues of affected piglets from a pig farm in Nanjing, Jiangsu, my country were collected as toxic materials. Put the toxic material in a mortar for aseptic operation first, then add 5ml of sterilized physiological saline, and grind it repeatedly; add 200U of penicillin and 200U of streptomycin to each ml of the treated toxic material, mix well, and mix it Put into -20 ℃ refrigerator and freeze and thaw repeatedly 3 times, freeze and thaw quickly each time to release the virus from the broken cells; centrifuge at 12000 r / min for 10min, take the supernatant and inoculate it into BHK21 cells (purchased from ATCC, That is, the American Type Culture Collection) culture. After 2 days of culture, with normal BHK21 cells ( figure 2 ) compared with that of BHK21 cells inoculated with toxic materials showed obvi...
Embodiment 2
[0056] Example 2 Preparation of vaccine composition containing inactivated porcine pseudorabies virus PRV-JS strain
[0057] 1. BHK21 cell seed propagation and expansion culture
[0058] Take out the frozen BHK21 cell tube from the liquid nitrogen tank, put it in a 37°C water bath to thaw quickly, transfer the cells into a centrifuge tube containing 10ml of DMEM medium, and centrifuge at 1000rpm for 5 minutes. Discard the supernatant, suspend the cells with the growth medium, then add to the cell culture flask, at 37°C, 5% CO 2 cultivated under conditions. When the cell coverage reached 100%, the cells were digested with 0.1% trypsin-EDTA solution. Subculture at a volume ratio of 1:3. The growth medium is DMEM solution containing 10% FBS (newborn bovine serum).
[0059] 2. Virus culture
[0060] The BHK21 cell culture with a cell growth coverage rate of 80% was removed from the growth medium. Inoculate PRV-JS strain virus solution according to the multiplicity of infecti...
Embodiment 3
[0070] Example 3 Safety inspection of porcine pseudorabies inactivated vaccine
[0071] Check the safety of the porcine pseudorabies inactivated vaccine prepared according to the method of Example 2.
[0072] Each batch of vaccines uses 30-35 day-old healthy piglets and healthy pregnant sows negative for pseudorabies antibodies, and each head is intramuscularly injected with 2 parts (4ml) of porcine pseudorabies inactivated vaccine, and another set of the same type that is not vaccinated Pigs were used as control group.
[0073] Observed for 14 days after inoculation, all experimental pigs observed indicators including body temperature, energy, appetite and other local and systemic reactions, the results are shown in Table 2. It can be seen from Table 2 that all test pigs showed no abnormality in spirit, body temperature, appetite, etc. The vaccinated piglets had no abnormal reaction caused by vaccination; the vaccinated pregnant sows had normal pregnancy and no abortion occ...
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