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A technology for fibroblasts and bursal disease, applied in the directions of viral antigen components, antiviral agents, viruses/phages, etc., can solve the problems of high cost, quality difference between product batches, chicken-derived potential disease infection, etc.
Active Publication Date: 2015-04-01
哈药集团生物疫苗有限公司
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Problems solved by technology
At present, the vast majority of commercially available bursal vaccines are chicken embryo tissue vaccines. Compared with cell vaccines, they have disadvantages such as high cost, large difference in product quality between batches, and easy infection of chicken-derived potential diseases.
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Embodiment 1
[0054] Example 1 Isolation of Infectious Bursal Virus Chicken Embryo Fibroblast Adapted Strain H11 Strain
[0055] 1. Virus isolation:
[0056] The disease material came from a farm in the suburbs of Acheng City, Heilongjiang Province. Dying bursa of Fabricius was collected, cut into pieces under aseptic conditions, treated with chloroform, centrifuged, and the supernatant was taken to inoculate chicken embryos, and chicken embryos and allantoic membranes were collected 48h-120h. After grinding with a tissue grinder, store at -20°C for later use.
[0057] 2. Preliminary identification of virus isolates
[0058] The hemagglutination test of the interstitial fluid was negative; the agar two-way diffusion test with rabbit anti-bursal virus polyclonal antibody was positive.
[0059]3. Passaging and harvesting on primary chicken embryo fibroblasts (CEF)
[0060] (1) Dilute the harvested tissue virus with sterilized saline, inoculate it on the CEF that grows into a monolayer, cul...
Embodiment 2
[0063] Example 2 Production of IBDV H11 Live Vaccine Using Passage Chicken Embryo Fibroblasts
[0064] 1. Preparation of poisonous seeds for production
[0065] (1) Breeding of poisonous species
[0066] The virus seed H11 strain (preservation number is CGMCC NO.6910) was prepared with sterilized physiological saline for 10 3 ~10 4 After one-fold dilution, a monolayer of chicken embryo fibroblasts (CEF) was inoculated. Discard the cell culture medium, inoculate according to 1 / 10 of the volume of the culture medium, absorb at 37°C for 1 hour, add the culture medium, and add 1% volume of 10% reduced glutathione solution to it. 37°C, 5% CO 2 conditions, cultured for 3 to 4 days, and harvested the virus liquid when 80% of the cells showed specific lesions. Freeze and thaw three times in a freezer at -20°C, harvest in a sterilized container, and store at -20°C. Indicate the date of harvest, the generation of the poisonous seeds and the loading amount.
[0067] (2) Identifica...
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Abstract
The invention relates to an infectious bursal disease virus chick embryo fibroblast adapted strain H11 and an application thereof. The preservation number of the infectious bursal disease virus chick embryo fibroblast adapted strain H11 is CGMCC NO. 6910, and the infectious bursal disease virus chick embryo fibroblast adapted strain H11 is preserved in the China General Microbiological Culture Collection Center (CGMCC). Besides, the invention also provides a method for separating H11 strain, and an application of the strain in preparing live vaccines for preventing an infectious bursal disease. The infectious bursal disease virus chick embryo fibroblast adapted strain H11 overcomes the defects that the cost of chick embryo tissue vaccines is high, the batch qualities of a product are in great difference, infections of chicken-derived potential diseases can be generated easily, and the like, so that the infectious bursal disease virus chick embryo fibroblast adapted strain H11 has extensive application prospects.
Description
technical field [0001] The invention belongs to the field of biological vaccines, and in particular relates to an infectious bursal virus chicken embryo fibroblast-adapted strain H11 and an application thereof. Background technique [0002] Chicken infectious bursal disease (Infeetious Bursal Disease, IBD) is a highly contagious disease caused by infectious bursal virus (Infeetious Bursal Disease Virus, IBDV). One of the diseases, mainly infringes on the bursa organs of chicks and young chickens. The harm of the disease not only directly causes the death of chickens, increases the elimination rate, affects the weight gain, but more importantly, destroys the B lymphocytes in the bursa, resulting in Immunosuppression makes sick chickens more susceptible to other diseases, and the immune response to vaccination is reduced or lost. [0003] Because IBD is relatively stable in the external environment, it is not easy to achieve the goal by taking disinfection and isolation measu...
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