Duck hemorrhagic ovaritis virus strain, inactivated vaccine and preparation method thereof
A technology for hemorrhagic oophoritis and inactivated vaccines is applied in the field of virus strains and inactivated vaccines, duck hemorrhagic oophoritis virus strains, inactivated vaccines and their preparation fields, which can solve the problem that the vaccine evaluation model has not been established and restricts vaccines. development and other issues
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Embodiment 1
[0038] a. Vaccination: the duck hemorrhagic oophoritis virus through the allantoic cavity according to 10 3 ELD 50 Inoculate 10-day-old sensitive duck embryos at a dose of 1 piece, and incubate at 37°C;
[0039] b. Harvesting: Harvest duck embryo bodies, chorioallantoic membranes, and allantoic fluid that died 48 to 120 hours after inoculation and had obvious disease scars, crushed at 20,000 r / min for 2 minutes, repeatedly frozen and thawed twice, and stored at -20°C. It is duck embryo tissue fluid; according to the method for determining the median lethal dose and the half infection dose specified in appendix 7 of "The Veterinary Pharmacopoeia of the People's Republic of China" (version three in 2010), the virus content assay is carried out, and the proliferating virus titer is 10 7.1 ELD 50 / 0.1ml;
[0040] c. Inactivation: Add formaldehyde solution with a mass concentration of 37% to the duck embryo tissue fluid to make the final content 0.2%, shake well and inactivate a...
Embodiment 2
[0049] a. inoculation: the duck hemorrhagic oophoritis virus cell adaptation virus is inoculated in the monolayer of duck embryo fibroblasts in a ratio of 0.1%;
[0050] b. Harvest: Harvest the cells when the cytopathic effect (CPE) reaches 80% to 90% 72 to 96 hours after inoculation, and put the sterile beaker containing the harvested cell pellet in an ice bath at 0°C with a cell lyser at 85 Under the condition of % power, lyse for 30s / time, lyse for 3 times, and freeze at -20°C to be the virus cell lysate; according to the median lethal dose, The virus content determination is carried out by the determination method of the half infection amount, and the proliferating virus titer is 10 6.5 TCID 50 / 0.1ml;
[0051] c. Inactivation: Add formaldehyde solution with a mass concentration of 37% to the virus cell lysate to make the final content reach 0.1%, shake well and inactivate at 4°C for 7 days;
[0052] d emulsification:
[0053] Preparation of aqueous phase: Centrifuge t...
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