A kind of canine parvovirus inactivated vaccine and preparation method thereof
A canine parvovirus and inactivated vaccine technology, which is applied in the field of canine parvovirus inactivated vaccine and its preparation, can solve the problem of decreased immune effect of inactivated vaccine and attenuated vaccine, genetic variation, and can not fully meet the needs of disease prevention and control and market consumption and other problems, to achieve the effect of high antibody production level, high protection level and good immunogenicity
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Embodiment 1
[0028] Example 1: Isolation and Identification of Canine Parvovirus CPV-HuN1703 Strain
[0029] 1.1 Source and collection of disease materials: The disease materials were collected from severely ill dogs admitted to a pet hospital in Hengyang City, Hunan Province, after death.
[0030] It was clinically found that the dog showed depression, lack of appetite, severe diarrhea accompanied by vomiting, red blood clots in the feces, and intestinal mucosal shedding. It was difficult to prevent the deterioration of the condition with existing drug means. The on-site loose stool was taken, and the CPV antigen was detected with a colloidal gold test strip, and the test result was positive.
[0031] After the dead dogs were disinfected, the heart, liver, spleen, lymph nodes, small intestine and intestinal contents of the dogs were collected and stored at -70°C.
[0032] Other sources of viruses:
[0033] CPV standard strain (CVCC AV298), from China Veterinary Drug Control (HA titer 1:...
Embodiment 2
[0054] Embodiment 2: Canine parvovirus CPV-HuN1703 strain is used for the preparation of vaccine
[0055] (1) Preparation of vaccine
[0056] An inactivated canine parvovirus vaccine, which is prepared by the following method:
[0057] Step a: Cultivate F81 feline kidney passage cells according to the conventional method: the growth medium is MEM containing 5% fetal bovine serum (containing penicillin sodium and streptomycin sulfate each 100 μg / mL), at 37°C, 5% CO 2 Cultured and subcultured in an incubator, and when the cells grew to 80% confluence, they were washed once with PBS and then replaced with serum-free medium;
[0058] Step b, take the content as 5×10 4 TCID 50 / ml of canine parvovirus CPV-HuN1703 strain was inoculated into the well-growing F81 cells prepared in step a at a volume percentage of 5%, adsorbed at 37°C for 1 hour, discarded the adsorption solution, and added MEM containing 10 μg / mL trypsin As a maintenance medium to continue culturing;
[0059] Ste...
Embodiment 3
[0068] Embodiment 3: Vaccine potency detection test
[0069] 3.1 Materials and methods
[0070] As a control, the CPV standard strain and the CPV-NY130615 strain were respectively used to prepare a batch of products according to the preparation method of the inactivated vaccine in Example 2. The commercialized vaccine uses Chongbiwei® Youmiankang (mainly immune to canine distemper, canine parvovirus, hepatitis and canine parainfluenza, batch number: A505A01)
[0071] Twenty-five 30-40-day-old hybrid puppies that were negative in both pathogen detection and antibody detection were randomly divided into 5 groups of 5 dogs, named as the control group, CPV-HuN1703 group, CPV standard group, CPV-HuN1703 group, and CPV-HuN1703 group, respectively. NY130615 group and commercial vaccine group, in which the control group was injected with 50% ISA206 adjuvant (50% ISA206+50% normal saline), and the CPV-HuN1703 group, CPV standard group, and CPV-NY130615 group were respectively injected...
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