Porcine parvnvirus living vaccine and preparation method thereof
A parvovirus, live vaccine technology, applied in the direction of antiviral agents, pharmaceutical formulations, medical preparations containing active ingredients, etc., can solve the problem of no effective treatment method for PPV infection, reduce cost burden, high antibody titer, Long shelf life effect
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Embodiment 1
[0035] A preparation method of porcine parvovirus disease live vaccine, the method comprises the following steps:
[0036] (1) Prepare ST cells according to conventional methods, the growth medium is MEM containing 8% newborn bovine serum, and when the cells grow into more than 2 / 3 monolayer, they are inoculated with poison;
[0037] (2) Take the attenuated porcine parvovirus PPV for production S-1 A strain virus (PPV S-1 The attenuated strain obtained by continuous subculture on the pig testicular cell line to the 40th generation or more, and its HA titer ≥ 2 9 , TCID 50 ≥107.0 / ml) was diluted at 0.1%, inoculated with a well-growing ST cell monolayer, adsorbed at 37°C for 1.5 hours, added MEM cell maintenance solution containing 2% newborn bovine serum, and placed at 37°C for static or rotational culture;
[0038] (3) Observing twice a day, recording the cytopathic condition, when more than 75% of the cells appear cytopathic, harvest the cell culture, freeze and thaw 3 ti...
Embodiment 2
[0058] A preparation method of porcine parvovirus disease live vaccine, the method comprises the following steps:
[0059] (1) Prepare ST cells according to conventional methods, the growth medium is MEM containing 5% newborn bovine serum, and inoculate when the cells grow into more than 2 / 3 monolayer;
[0060] (2) Take the attenuated porcine parvovirus PPV for production S-1 A strain virus (PPV S-1 The attenuated strain obtained by continuous subculture on the pig testicular cell line to the 40th generation or more, and its HA titer ≥ 2 9 , TCID 50 ≥10 7.0 / ml) was diluted at 0.1%, inoculated with a well-growing ST cell monolayer, adsorbed at 37°C for 1.5 hours, added MEM cell maintenance solution containing 2% newborn bovine serum, and placed at 37°C for static or rotational culture;
[0061] (3) Observing twice a day, recording the cytopathic condition, when more than 75% of the cells appear cytopathic, harvest the cell culture, freeze and thaw 3 times, and store it bel...
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