Swine fever oral attenuated freezing-dry vaccine and preparation method thereof and freeze-drying protective agent
A technology of freeze-dried protective agent and attenuated vaccine, which is applied in the field of oral attenuated freeze-dried vaccine for swine fever and freeze-dried protective agent, can solve the problems of increasing the difficulty of screening freeze-dried protective agent, and avoid the shrinkage and activity reduction of freeze-dried products. loss of protective activity
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[0053] The preparation method and evaluation of oral attenuated freeze-dried vaccine for classical swine fever include:
[0054] 1. Primer design and synthesis
[0055] According to the CSFV E2 gene sequence, a pair of PCR detection primers were designed and synthesized. The size of the PCR product is 287bp. The specific sequence is as follows:
[0056] CSFV-F1:CAGGTATGCGATCTCGTCAACCA
[0057] CSFV-R1: GGGCACAGCCCAAATCCGAAGT
[0058] According to the mouse IFN-γ and IL-4 genes, a pair of fluorescent quantitative PCR primers for IFN-γ and IL-4 were designed and synthesized respectively. The primers were synthesized by Huada Gene Technology Service Company. The specific sequences are as follows:
[0059] IFN-γ-1: CCATCGGCTGACCTAGAGAA
[0060] IFN-γ-2: GATGCAGTGTGTAGCGTTCA
[0061] IL-4-1: GGTCTCAACCCCCAGCTAGT
[0062] IL-4-2: GCCGATGATCTCTCTCTCAAGTGAT
[0063] 2. Isolation and identification of CSFV
[0064] Use PK15 cells to proliferate CSFV. When the confluence of PK15 ...
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