Duck circovirus and adenovirus bivalent inactivated vaccine and preparation method of yolk antibody thereof
A dual inactivated vaccine and duck circovirus technology, which is applied in the field of poultry epidemic immunization control, can solve problems such as large market vacancies, achieve the effect of improving antibody titer, optimizing immunization procedures and extraction steps, and good protection effect
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Embodiment 1
[0052] The preparation of embodiment 1 vaccine
[0053] The original virus species of circovirus was isolated from the liver of ducks with clinical onset in 2019. PCR detection technology was used to confirm that the diseased ducks were infected with duck circovirus, and other pathogens were detected to rule out the possibility of virus mixed infection, and then homogenized and centrifuged. The supernatant was filtered and sterilized with a disposable filter; the virus seed infected 1-day-old ducklings, and was continuously passed down for 3 generations. After the virus amount was detected to be qualified, it was used as a purified virus seed. The inventor carried out biological preservation of the virus strain, and the preservation number: CGMCC No.19296;
[0054] Adenovirus (serotype IV) strains were purchased from China Veterinary Microbiology Culture Collection Center (preservation number: CVCCAV211);
[0055] The 1-day-old healthy Cherry Valley ducklings were selected to...
Embodiment 3
[0084] Embodiment 3 Vaccination dose and safety evaluation
[0085] According to the conclusion drawn in Example 2, according to duck circovirus and adenovirus antigen ratio 3:1, pine pollen polysaccharide concentration 20mg / mL, produce vaccine, and evaluate the vaccination dose and safety of vaccine. The vaccination dose was set as three doses of 0.2, 0.5, and 1.0 mL, and each dose was inoculated with 5 ducklings at the age of 1, 4, and 7 days (only once inoculated, no additional immunization). Venous blood was collected, serum was separated, and the titer of duck circovirus antibody was detected by agar diffusion test, and the result was expressed as the mean value ± SD of titer log2. The stress situation of each group after inoculation was recorded, and the stress manifested as depression, reluctance to move around, reduced food intake, etc., and death in severe cases.
[0086] The results in Table 3 show that the age of inoculation has little effect on antibody production...
Embodiment 4
[0092] The making of embodiment 4 yolk antibody
[0093] (1) Use the vaccine used in Example 3 to immunize the laying hens 1 month before the start of laying in 4 times, the inoculation dose is 1.5mL / only, and the interval between each immunization is 2 weeks, and the follow-up is every 1-2 months immune once;
[0094] (2) Collect immunized eggs two weeks after the fourth immunization, and keep immunizing once every 1-2 months during the continuous egg harvesting process; separate the egg yolk from the egg white with an egg yolk separator, and separate the egg yolk from the egg yolk preheated to 35°C. Dilute the egg yolk liquid with purified water at a volume ratio of 3.5:1, stir evenly to obtain the egg yolk dilution, and pre-warm at 35°C for 1 hour.
[0095] (3) Add PEG6000 with a final concentration of 3.5% to the egg yolk dilution, mix it with the egg yolk, stir well, and let it stand for 4 hours to fully react.
[0096] (4) Take out the supernatant after the reaction, a...
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