Porcine epidemic diarrhea virus strain and inactivated vaccine and interferon combined immunization method thereof
A technology of porcine epidemic diarrhea and interferon, which is applied in the field of viruses and immunity, can solve problems such as incompatibility, and achieve the effects of good immunogenicity, good immunogenicity and stability, and wide application value
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Embodiment 1
[0022] Example 1 Isolation, Identification and Sequencing Analysis of Porcine Epidemic Diarrhea Virus PEDV-CH-HB-1801 Strain
[0023] 1. Virus isolation and identification
[0024] Take the small intestine and its contents of piglets with suspected porcine epidemic diarrhea cases collected clinically, after aseptic treatment, grinding and crushing, centrifuge at 12000rpm at 4°C for 5 minutes, collect the upper layer suspension, filter through a 0.22um sterile filter, and collect filtrate. Extract RNA, use the cDNA obtained by reverse transcription as a template, and use the primers shown in SEQ ID NO.1 to amplify the target fragment of the virus. Agarose gel electrophoresis was used to identify the size of the target fragment and determine whether it was a popular strain of PEDV.
[0025] The filtrate was taken and inoculated into a monolayer of Vero cells at a ratio of 500ul per well of a 6-well plate. At the same time, an appropriate amount of trypsin was added to the cel...
Embodiment 2
[0046] Example 2 Establishment of the joint immunization method of porcine epidemic diarrhea virus PEDV-CH-HB-1801 strain and porcine genetically engineered interferon
[0047] According to the method described above, the inactivated vaccine of PEDV-CH-HB-1801 strain was prepared. At the same time, the porcine genetically engineered interferon diluted in physiological saline or astragalus polysaccharide injection was prepared respectively. The concentration of astragalus polysaccharide in the astragalus polysaccharide injection is 10 mg / mL. The porcine genetically engineered interferon is alpha interferon, and the final dilution concentration is 500,000 IU / mL.
[0048] Fifteen PEDV-negative pregnant sows were selected and divided into 5 groups with 3 pigs in each group. At 40 days and 20 days before delivery, the sows were injected with immune inactivated vaccine and pig genetically engineered interferon once in each Houhai point. The grouping situation is shown in Table 4....
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