Infectious hematopoietic organ necrosis vaccine and method for amplifying virus of infectious hematopoietic organ necrosis vaccine on muscle cells of Pimephales promelas
A technology for necrosis of hematopoietic organs and muscle cells, applied in the biological field, can solve the problems of prolonging the production cycle, increasing the probability of pollution, wrong packaging, etc., and achieve the effect of stable virus titer, high virus titer and good immune effect
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Embodiment 1
[0031] The preparation of embodiment 1, IHNV virus liquid
[0032] 1. Recovery and culture of FHM cells
[0033] Take out the FHM (American Type Culture Collection, American Type Culture Collection, No.: CCL-42 TM ), quickly put it into a 30°C constant temperature water bath, and shake it from time to time to completely melt it. Move it into a 15ml centrifuge tube under aseptic conditions, centrifuge at 1000g for 2min, discard the supernatant, add 1ml of cell culture medium (M199 medium + volume percentage 10% fetal bovine serum + mass volume percentage (g: ml) 1% double antibody) Gently blow evenly. Transfer the blown cell solution into a T-25 cell culture bottle, add 4ml of cell culture solution, blow it evenly, put it in a 25°C carbon dioxide constant temperature cell incubator for static culture, and observe the cell state under an inverted microscope every day. When the T-25 cell culture flask is covered with a monolayer of FHM cells, digest the adherent cells with 1...
Embodiment 2
[0065] Preparation and Application of Embodiment 2, IHNV Inactivated Vaccine
[0066] One, the preparation of IHNV inactivated vaccine
[0067] IHN-BPL inactivated vaccine: add β-propiolactone (BPL) to the amplified IHNV virus liquid obtained in 3 of Example 1 of 80ml, so that the final concentration of BPL is 3mM, mix and place on a shaker at 24°C , 100r / min carried out inactivation for 24h, adding sodium thiosulfate solution (final concentration is 20mM) to terminate the inactivation, to obtain IHN-BPL inactivated seedlings;
[0068] IHN-formaldehyde inactivated vaccine: add formaldehyde to 80ml of the amplified IHNV virus liquid obtained in 3 of Example 1, so that the final concentration of formaldehyde is 5mM, mix well, place on a shaker at 24°C, and inactivate at 100r / min After 24 hours, sodium bisulfite solution (final concentration: 1 mM) was added to terminate the inactivation to obtain IHN-formaldehyde inactivated vaccine.
[0069] 2. Detection of relative immune pr...
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