Cell inactivated vaccine, egg yolk antibody injection, and preparation method of cell inactivated vaccine
A technology of inactivated vaccine and yolk antibody, which is applied in the field of cell inactivated vaccine, yolk antibody injection and preparation thereof, can solve problems such as labor-intensive and time-consuming, and achieve the effect of reducing production steps, achieving remarkable curative effect, and reducing the probability of morbidity.
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Embodiment 1
[0025] Example 1: Preparation of porcine circovirus cell inactivated vaccine in this example. The disease material was collected from pig farms with severe porcine circovirus disease in Shandong area. After clearing and filtering through a 0.22 μm filter membrane, a sensitive monolayer of PK-15 cells was inoculated. After the PK-15 cells were continuously passaged for 6 generations, part of the cells were frozen and thawed to extract the DNA of the PCV-2 virus, and the target gene was amplified with PCR primers specific to the PCV-2 virus ORF1 gene. The amplified product was cloned into the PMD-18T vector for sequencing, identified and named SD strain. Multiple viral epitopes with antigenic immune activity were screened out in this strain, and five antigenic epitopes were finally selected after specific primer PCR, product electrophoresis, sequencing, cloning, purification, and detection of protein immune activity of each epitope. site, each antigen site is connected by splic...
Embodiment 2
[0047] Embodiment 2: The preparation of porcine circovirus cell inactivated vaccine of the present embodiment collects disease material from pig farm that serious porcine circovirus takes place in Shandong area, and disease material is after homogenization, freeze-thaw repeatedly 3 times, high-speed centrifugation, take After the supernatant was filtered through a 0.22 μm filter membrane, a sensitive monolayer of PK-15 cells was inoculated. After the PK-15 cells were continuously passaged for 6 generations, part of the cells were frozen and thawed to extract the DNA of the PCV-2 virus, and the target gene was amplified with PCR primers specific to the PCV-2 virus ORF1 gene. The amplified product was cloned into the PMD-18T vector for sequencing, identified and named SD strain. Multiple viral epitopes with antigenic immune activity were screened out in this strain, and five antigenic epitopes were finally selected after specific primer PCR, product electrophoresis, sequencing, ...
Embodiment 3
[0069] Embodiment 3: The preparation of porcine circovirus cell inactivated vaccine of this embodiment collects disease material from pig farm that serious porcine circovirus takes place in Shandong area, and disease material is after homogenization, freeze-thaw repeatedly 3 times, high-speed centrifugation, take After the supernatant was filtered through a 0.22 μm filter membrane, a sensitive monolayer of PK-15 cells was inoculated. After the PK-15 cells were continuously passaged for 6 generations, part of the cells were frozen and thawed to extract the DNA of the PCV-2 virus, and the target gene was amplified with PCR primers specific to the PCV-2 virus ORF1 gene. The amplified product was cloned into the PMD-18T vector for sequencing, identified and named SD strain. Multiple viral epitopes with antigenic immune activity were screened out in this strain, and five antigenic epitopes were finally selected after specific primer PCR, product electrophoresis, sequencing, cloning...
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