Dog parvovirus attenuated vaccine strain and application thereof
A technology of canine parvovirus and attenuated vaccine, which is applied in the field of preparing drugs for preventing or treating diseases caused by canine parvovirus and virus attenuated vaccine strains, can solve the problems of high cost and weak antigen targeting, and achieves good effects, Prevention or treatment of canine distemper, good protective effect
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Embodiment 1
[0012] Example 1 Isolation and identification of canine parvovirus virulent isolate (CPV-YN)
[0013] 1 Materials and methods
[0014] 1.1 Materials
[0015] 1.1.1 Disease material The diseased and dead dogs of Yinong Comprehensive Farm in Daowai District, Harbin City.
[0016] 1.1.2 CRFK cells (65-95 generations the same below) were purchased from China Veterinary Drug Control Institute and cultured according to conventional methods.
[0017] 1.1.3 As experimental animals, puppies aged 2 to 4 months (CPV antibody titer ≤ 1:2) were purchased from the Department of Experimental Zoology of Harbin Medical University.
[0018] 1.1.4 Serum CPV standard positive serum (antibody titer ≥ 1:16) and CPV negative serum were provided by Harbin Veterinary Research Institute.
[0019] 1.2 Separation method
[0020] 1.2.1 Isolation of virus 5 samples of bloody feces from sick dogs were prepared into a 1:9 suspension with MEM cell nutrient solution, mixed evenly, centrifuged at 5000r / min ...
Embodiment 2
[0066] Embodiment 2 The cultivation of canine parvovirus attenuated vaccine strain CPV-YNR
[0067] 1.1 Test material
[0068] The test animals are healthy dogs (CPV antibody titer ≤ 1:2) of 2 to 4 months of age; anti-canine parvovirus-specific serum and 0.2% erythrocyte suspension are prepared by our laboratory, and the virulent is isolated from Example 1. CPV-YN 6 generations.
[0069] 1.2 Cultivation and identification of CPV-YN attenuated strain
[0070] 1.2.1 Cultivation of CPV-YN attenuated strain Passage CPV-YN to 110 generations through CRFK cells.
[0071] 1.2.2 Morphological observation of attenuated CPV-YN strain
[0072] Centrifuge the harvested CPV-YN 5, 10, 20, 30, 50, 70, 90, 100, 105, and 110 passage virus cell cultures at 5000r / min for 30min, take the supernatant and ultracentrifuge at 20000rpm, and centrifuge at 4°C for 2h. Suspend the precipitate with an appropriate amount of PBS (pH 7.0), and observe with a negative staining electron microscope with pho...
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