Applications of porcine reproductive and respiratory syndrome virus (PRRSV), mycoplasma hyopneumoniae and PCV-2 antigen in preparation of vaccine
A technology of mycoplasma hyopneumoniae and PCV-2, applied in bacterial antigenic components, viral antigenic components, antibacterial drugs, etc., can solve problems such as delaying PRRSV vaccine immunization time, reducing pig herd tolerance to PRRSV, prolonging piglet immunization time of PRRSV vaccine, etc.
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Embodiment 1
[0034] Embodiment 1, the preparation of porcine circovirus type 2, PRRSV, mycoplasma hyopneumoniae combination vaccine
[0035] 1. Antigen Preparation
[0036] 1.1 Preparation of porcine circovirus type 2 antigen
[0037] (1) Passaging and culturing of cells for seedling production: PK15 cells were taken, digested and passaged by EDTA-trypsin cell dispersion solution (Hank's solution containing 0.25 trypsin (1:250), 0.02% EDTA), to obtain 90% MEM solution, 10% bovine serum, each 100 units / ml of penicillin sodium and streptomycin sulfate, the cell growth medium adjusted to pH 7.2 to continue culture, culture temperature is 37 ℃, when a good cell monolayer is formed, it is used for continued passage Or inoculated in a bioreactor for microcarrier suspension culture, wherein the bioreactor is a bioreactor that can automatically control parameters such as temperature, pH, dissolved oxygen, and stirring speed, and is suitable for microcarrier suspension culture.
[0038] (2) Propa...
Embodiment 2
[0058] Embodiment 2, PCV-2, PRRSV, mycoplasma hyopneumoniae combination vaccine application
[0059] Use the self-made component I and component II in Example 1 of the present invention in combination, and use component I as the diluent to dilute component II to form a joint vaccine to prevent the three diseases of Mycoplasma hyopneumoniae, PCV-2 and PRRSV. The immunization procedure is as follows: Firstly, 7-day-old piglets are immunized with component I diluted with component II for the first vaccination, and 21 days later, component I is used for the second immunization. At the same time of immunization, the control group was immunized with the corresponding single vaccine to evaluate the difference in immune effect between the combined application of the three antigen components and the single use of the corresponding single antigen.
[0060] 1. Materials
[0061] PCV-2 and Mycoplasma hyopneumoniae dual vaccine, ie component I, was prepared and tested according to Example...
Embodiment 3
[0092] Example 3, Application of PCV-2 Antigen, Mycoplasma Hyopneumoniae Antigen and PRRSV Attenuated Strain Combination Vaccine in Pigs Infected with Mycoplasma Hyopneumoniae
[0093] 1. Test method
[0094] 1.1 Research purpose and experimental design
[0095] In Example 2, the test animals selected for the evaluation of the immune effect of the multi-component vaccine and the single vaccine were antigen-antibody double-negative piglets, but Mycoplasma hyopneumoniae widely exists in pig herds in actual clinical practice. Therefore, this embodiment aims at immunizing porcine PCV-2, PRRSV and Mycoplasma hyopneumoniae in pig herds with Mycoplasma hyopneumoniae in order to evaluate the immune effect. This example included piglets from 3 scale farms with M. hyopneumoniae, each farm containing about 200 piglets. The piglets in each pig farm were divided into two groups: the first group of 7-day-old piglets were intramuscularly injected with the triple vaccine consisting of compo...
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