Vaccine composition containing swine mycoplasmal pneumonia antigen and swine streptococcosis antigen, and preparation method and application thereof
A technology of mycoplasma pneumonia and vaccine composition, which is applied in the direction of antibacterial drugs and bacterial antigen components, which can solve the problems of drug resistance, stress and safety problems, residues, etc., to reduce the number of vaccinations, enhance the immune effect and effect better effect
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Embodiment 1
[0038] Embodiment 1, preparation of Mycoplasma hyopneumoniae antigen, Streptococcus suis antigen
[0039] 1. Source of bacteria (virus) strains
[0040] The selected Mycoplasma hyopneumoniae (Mhp for short) is HN0613 strain, the preservation number is CCTCCM2012230, and the preservation date is June 13, 2012.
[0041] The selected Streptococcus suis Type 2 (SS2 for short) is an SC strain, the preservation number is: CCTCCM2011351, and the preservation date is October 13, 2011.
[0042] The selected strain of S. equi subsp. zooepidemicus (SESZ for short) is ATCC35246 strain (purchased from American type culture collection, namely American type culture collection, ATCC)
[0043] 2. Preparation and inspection of vaccine semi-finished products
[0044] 2.1 Preparation of seeds for production
[0045] 2.1.1 Streptococcus suis type 2, Streptococcus equi zooepidemic subspecies:
[0046] Propagation of primary seeds
[0047] Dilute the SS2SC strain and SESZ ATCC35246 freeze-dri...
Embodiment 2
[0073] Example 2. Preparation of vaccine composition
[0074] 1 Preparation of preservatives
[0075] 1% (w / v) thimerosal (Sinopharm Chemical Reagent Co., Ltd. 20120106) aqueous solution
[0076] 2 preparation of diluent
[0077] Sterile PBS (0.01M, pH=0.02) buffer solution: dissolve 8g sodium chloride, 0.25g potassium chloride, 3.63g disodium hydrogen phosphate, 0.24g potassium dihydrogen phosphate in 900ml purified water, then dilute to 1L, autoclave at 121°C for 30 minutes for later use.
[0078] 3 preparation of adjuvants
[0079] Gel 01 (French SEPPIC company 20120401), autoclaved at 121°C for 30 minutes for later use.
[0080] 4 with seedlings
[0081] Get the concentrated inactivated antigen of Mhp, SS2, SESZ prepared in Example 1, make mixed antigen solution or directly make antigen solution according to the final antigen content in combined vaccine and single vaccine, then antigen solution and Gel01 adjuvant by 90: 10 (v / v) mixed, supplemented to the required vo...
Embodiment 3
[0106] Example 3. This study is to evaluate the combination efficacy of Mhp-SS2-SESZ combination vaccines with different antigen contents.
[0107] 1 Material:
[0108] Embodiment 2 prepares vaccine:
[0109] Table 12
[0110]
[0111]
[0112] Animals: 4-5 weeks old, pigs with negative serum antibodies to Mhp, SS2, and SESZ
[0113] Challenge strains: SS2SC strain, SESZ ATCC35246 strain (purchased from American type culture collection, ATCC), Mhp CVCC354 strain (purchased from China Veterinary Drug Control Institute)
[0114] 2 methods:
[0115] 50 piglets aged 4-5 weeks were selected and randomly divided into 10 groups, 5 piglets in each group. On the 0th day, each pig in groups 1, 4, and 7 was injected with vaccine L intramuscularly in the neck, and each pig in groups 2, 5, and 8 was injected with vaccine H in the neck. Intramuscular injection of sterile PBS, both 2ml / head, the 10th group was used as blank control, neither immunized nor challenged. The 1st to 6t...
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