Vaccine composition, and preparation method and application thereof
A technology of vaccine composition and swine fever virus, which is applied in the field of veterinary biological products, can solve problems such as economic loss and immunization failure of swine fever vaccine
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Embodiment 1
[0050] Preparation of Antigens of Swine Fever and Streptococcus Suis
[0051] 1 Preparation of swine fever antigen
[0052] 1.1 Digest and disperse the highly sensitive ST cells (purchased from ATCC) that have grown into a good monolayer and are highly sensitive to the attenuated rabbit strain of CSFV with a digestive solution containing 0.125% trypsin and 0.03% EDTA, and inoculate the cells for cell culture after counting the cells bottle, add 3% calf serum MEM cell culture solution, and at the same time add seed poison according to M.O.I.=0.1 inoculation dose, and place it in a 37°C incubator for cultivation.
[0053] After cultivating for three days, the poison was collected for the first time, and the cell maintenance solution containing 1.5% calf serum was added after the poison collection, and the poison was collected once every 2 days thereafter, and the poison could be collected 5 times continuously. After harvesting the virus, the antigens were mixed and stored at -2...
Embodiment 2
[0086] Preparation of swine fever and Streptococcus suis vaccine composition
[0087] 1 Combined use of inactivated vaccine diluted with live vaccine
[0088] 1.1 Get the hog fever whole virus antigen prepared in Example 1, add heat-resistant protective agent (2wt% gelatin aqueous solution and 15wt% lactose aqueous solution are prepared in 1:1 (v / v) ratio) with 1:1 (v / v) ratio After mixing, fully mix, quantitatively dispense, and freeze and vacuum-dry quickly, that is, live swine fever vaccine.
[0089] 1.2 Take the whole bacterial antigen of Streptococcus suis prepared in Example 1, slowly add it to the water-soluble adjuvant gel adjuvant (Seppic, France), and continuously stir it with an emulsifier at 800 rpm for 12 minutes during the addition process, and mix well , stored at 4°C, it is the inactivated vaccine of Streptococcus suis.
[0090] 1.3 When in use, dilute the live swine fever vaccine with inactivated Streptococcus suis vaccine, and the specific formulation of th...
Embodiment 3
[0099] Immunogenicity test of swine fever and Streptococcus suis vaccine composition
[0100] 1 Immunization program for piglets
[0101] 170 piglets that were 25-30 days old and negative for swine fever and Streptococcus suis antigen antibodies were divided into 17 groups for the test. The 1st-2nd group was the vaccine 1 immunization group, the 3rd-4th group was the vaccine 2 immunization group, and the 5th- Group 6 is vaccine 3 immunization group, Group 7-8 is vaccine 4 immunization group, Group 9-10 is vaccine 5 immunization group, Group 11-12 is vaccine 6 immunization group, and Group 13-14 is vaccine 7 immunization group group, the 15th-17th group is the PBS control group. Each pig in the PBS control group was injected with 1ml of PBS with a ratio of 10% (V / V) gel adjuvant, and each pig in the vaccine immunized group was injected with 1ml of the vaccine in the neck muscle for a single immunization.
[0102] 2 Challenge protection test of immune piglets
[0103] 21 days...
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