Porcine seneca valley virus vaccine and preparation method thereof
A virus vaccine, virus technology, applied in biochemical equipment and methods, viruses, vaccines, etc., can solve problems such as slow weight gain, economic loss, high mortality, etc.
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Embodiment 1
[0065] Embodiment 1——The gene characteristic of SVV-SD-2018 strain ORF fragment
[0066] The structural protein of the SVV-SD-2018 strain is the ORF region. The ORF fragment was amplified and sequenced by PCR using 5 pairs of primers. The homology comparison showed that the amino acid sequence of this fragment was compared with domestic reference strains / epidemic strains. Mutations occurred at positions 165, 495, 575, 770, 1119, 1476, 1609, 1854, 2065, 2104 and 2109; , 1119, 1469, 1476, 1609, 1854, 2065, 2104 and 2109 were mutated.
Embodiment 2
[0067] Embodiment 2——Preparation of poisonous seeds for production
[0068] Add the SVV-SD-2018 virus seed into the well-growing BHK-21 cell suspension culture at the ratio of MOI=0.01~0.5, culture at 37°C, harvest when the cell viability rate is lower than 50%, quantitatively aliquot, and cryopreserve. Indicate the date of harvest, algebra of poisonous species, etc. The virus content remained stable at 10 9.5 TCID 50 / mL or more.
Embodiment 3
[0069] Embodiment 3——Preparation of Porcine Seneca Virus Inactivated Vaccine
[0070] 1. Cell Culture and Inoculation
[0071] (1) Shake flask culture process
[0072] 1) Effects of different multiplicity of infection on virus content The initial cell density was 2.5×10 6 Cells / mL BHK-21 cells were inoculated with SVV-SD-2018 strain at MOI of 0.01, 0.05, 0.1, and 0.5, respectively, and samples were taken 24 hours, 36 hours, and 48 hours after inoculation to determine the virus content.
[0073] 2) Effect of cell density on virus content when inoculated BHK-21 cells were inoculated into shake flasks, when the cell density was 2.0×10 6 cells / mL, 2.5×10 6 cells / ml, 3.0×10 6 cells / mL, 3.5×10 6 cells / mL, 4.0×10 6 Cells / mL, the SVV-SD-2018 strain was inoculated according to MOI=0.05, samples were taken 24 hours, 36 hours, and 48 hours after inoculation, and the virus content was determined.
[0074] 3) Effect of different harvesting times on virus content Inoculate shake flas...
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