Combined live vaccine against porcine reproductive and respiratory syndrome, swine fever and pseudorabies, and preparation method thereof
A technology for respiratory syndrome and pseudorabies, which can be used in medical preparations, antiviral agents, and pharmaceutical formulations containing active ingredients, which can solve problems such as being difficult to achieve, affecting the effect of disease prevention, and time-consuming and labor-intensive.
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[0065] In some embodiments, the preparation method may include: (1) subculture and culture the susceptible cell line; (2) the highly pathogenic porcine reproductive and respiratory syndrome TJM strain, the attenuated strain of classical swine fever rabbit The C strain and the pseudorabies Bartha K61 strain were inoculated with susceptible cell lines respectively, and were cultured with a maintenance solution to prepare production virus seeds; (3) the prepared production virus seeds were respectively inoculated on mediums full of 90-100% cells, and maintained with The highly pathogenic porcine reproductive and respiratory syndrome TJM strain virus antigen solution, the classical swine fever lathe attenuated strain cytotoxic antigen solution and the pseudorabies Bartha K61 strain virus antigen solution were obtained by liquid culture and proliferation; (4) the highly pathogenic pig breeding Mix it with respiratory syndrome TJM strain antigen solution, swine fever lathe attenuated...
Embodiment 1
[0083] Embodiment 1 uses cell line to produce the method for highly pathogenic porcine reproductive and respiratory syndrome, swine fever and pseudorabies triple live vaccine
[0084] (1) Subculture and cultivation of cells for seedling production:
[0085] The highly pathogenic PRRSV TJM strain vaccine was prepared with Marc-145 cell line. The Marc-145 cell line was digested and passaged by trypsin-EDTA cell dispersion, and passaged one to three. Add cell growth solution and continue culturing at 37°C. When a good monolayer is formed, it is used for continued passage or virus inoculation.
[0086] The BT cell line was used to prepare the C-strain vaccine of the attenuated rabbit strain of CSF. The BT cell line was digested and passaged with trypsin-EDTA cell dispersion, and passaged at a ratio of 1 to 5; cell growth medium was added to continue culture at 37°C, and when a good monolayer was formed, it was used for continued passage or virus inoculation.
[0087] The vaccin...
Embodiment 2 3
[0096] The immunosuppressive test of embodiment 2 triple live vaccines
[0097] The test used 46 healthy piglets aged 21-28 days, and the test was divided into 10 groups. Among the healthy piglets used, the antigens and antibodies of highly pathogenic porcine reproductive and respiratory syndrome, classical swine fever and pseudorabies were all negative. The first group uses 5 test animals, and the single vaccine of the highly pathogenic PRRSV live vaccine TJM strain is injected intramuscularly into the neck, and the dose of the inoculated virus seed is 10 5.0 TCID 50 / ml, 1ml / pig. The second group uses 5 test animals, and the single vaccine of the C strain of classical swine fever rabbitization attenuated cytotoxic vaccine is injected intramuscularly into the neck. 4.0 TCID 50 / ml), 1ml / pig. The third group used 5 experimental animals, and intramuscularly injected a single vaccine of the Bartha K61 strain of the pseudorabies virus vaccine into the neck, and the dose of t...
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