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Preparation method of composite inactivated vaccine for IBD (infection bursal disease)

A technology for bursal disease and chicken infectivity, which is applied in the field of preparation of a compound inactivated vaccine for infectious bursal disease of chickens, can solve the problems of poor immune effect, high price, not being widely promoted, and the like, so as to improve immunity. , the effect of improving immunity and improving the ability to prevent diseases

Inactive Publication Date: 2016-03-30
浙江美保龙生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

According to the clinical use of live vaccines at home and abroad, although live vaccines can play a part of the protective effect, there is a shrinkage of the bursa of Fabricius itself, which leads to the disorder of the autoimmune system. In addition, genetic engineering vaccines, antigen-antibody complex vaccines There is also a low protection rate
Inactivated vaccines can produce higher antibodies after immunizing chickens, which can effectively protect the body. However, at present, domestic inactivated bursa vaccines have not been widely promoted due to their poor immune effect or high production costs. Most of the vaccines used are expensive imported vaccines

Method used

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  • Preparation method of composite inactivated vaccine for IBD (infection bursal disease)
  • Preparation method of composite inactivated vaccine for IBD (infection bursal disease)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Embodiment 1 chicken infectious bursal disease virus suspension preparation

[0034] (1) Subculture of DF-1 cells

[0035]Within 24-48 hours of culture, take the cell bottle with good shape, uniform distribution and full monolayer, discard the cell culture medium, add an appropriate amount of trypsin solution to the cell culture bottle for digestion; pour off the cell digestion solution, and then add the cell culture medium to the cell culture bottle. Add DMEM / F12 cell growth medium containing 10% fetal bovine serum into the bottle, gently blow the cell surface with a pipette to make the cells fall off and disperse evenly, and then add the cell growth medium at a ratio of 1:3-4 for passage.

[0036] (2) Inoculation

[0037] 24-48 hours after the cells grow into a single layer, change the medium and inoculate the virus. Take the virus species (Infectious Bursal Virus HQ strain) for production and inoculate DF- 1 cell and continue to culture at 37°C. The cell maintenan...

Embodiment 2

[0043] The preparation of embodiment 2 Chinese medicine flavonoids

[0044] a. Weigh 25 parts of chrysanthemum, 25 parts of raw land, 35 parts of vitex, 18 parts of gentian, 25 parts of scutellaria, 26 parts of Smilax smilax, and 25 parts of scrophulariaceae, chop them up, wash them and wash them with cold water Soak overnight, then add purified water 15 times the weight of raw materials, water bath to 90°C, and keep at 90°C, cook for 2 hours, stir every 10 minutes during the cooking process;

[0045] b. Discard the dregs of the traditional Chinese medicine in a, collect the medicinal liquid, and centrifuge at 10,000 rpm for 10 minutes after cooling at room temperature, discard the precipitate, and collect the supernatant;

[0046] c. The supernatant in b is subjected to alcohol precipitation, and the supernatant is obtained crude Chinese medicine flavonoids;

[0047] d. Concentrate the supernatant in c under reduced pressure and vacuum, dilute and wash the concentrate with s...

Embodiment 3

[0049] The preparation of embodiment 3 bursalin

[0050] a. Take healthy bursa tissue, remove the fascia and adipose tissue from the bursa tissue, and wash it with pH 7.2 sterilized cold PBS.

[0051] b. Weigh the bursa of Fabricius, add pH 7.2 sterilized cold PBS at a ratio of 1:1 (W:V), and perform high-speed homogenization in a tissue homogenizer.

[0052] c. Add 2.5% trypsin to the homogenate, freeze and thaw three times, centrifuge at 12000rpm for 20min, and discard the precipitate.

[0053] d. The supernatant is ultrafiltered with an ultrafiltration membrane with a molecular weight cut-off of 1000da, and the liquid under the membrane is the crude extract of bursalin.

[0054] e. The crude extract in d is sterilized by filtration through a 0.22um filter membrane, and then freeze-dried in a vacuum to obtain bursalin.

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Abstract

The invention discloses a preparation method of a composite inactivated vaccine for an IBD (infection bursal disease) and belongs to the technical field of animal biological products. The preparation method comprises technological steps as follows: 1), virus reproduction; 2), preparation of the composite inactivated vaccine for the IBD: 1-2 parts of an immunopotentiator is added to each 1,000 ml of an IBDV (infection bursal disease virus) antigen solution checked to be qualified in inactivation, the mixture is uniformly mixed, emulsification packaging is performed on the mixture and a summit livestock water-in-oil-in-water adjuvant in the volume ratio being 3:2, and the composite inactivated vaccine for the IBD is prepared. The novel vaccine adjuvant is adopted, and the prepared vaccine has double characteristics of water in oil and oil in water, combines the sustained release function of an ordinary vaccine and has advantages of joint actions of the water-soluble immunopotentiator, flavone of traditional Chinese medicines, a pain killer and the like. With the adoption of the effective immunopotentiator, the immunity of a body is greatly improved, the cellular immunity capability of the inactivated vaccine is improved, the body is stimulated to generate efficient neutralizing antibodies, and the disease prevention capability of the body is improved.

Description

technical field [0001] The invention belongs to the technical field of veterinary biological products, and in particular relates to a preparation method of a chicken infectious bursal disease compound inactivated vaccine. Background technique [0002] Chicken infectious bursal disease (Infection bursal disease, IBD) is a highly contagious infectious disease of chickens and turkeys caused by infectious bursal virus (Infection bursal disease virus, IBDV), mainly infecting young chickens aged 3 to 12 weeks , the virus mainly proliferates in the lymphocytes of the bursa of Fabricius, and causes varying degrees of damage to other immune organs, eventually leading to atrophy of the bursa of Fabricius in chickens and causing immunosuppression in chickens. The disease can clinically lead to symptoms such as weight loss, slow growth and skeletal muscle hemorrhage, and chickens less than 3 weeks old will not show obvious clinical features. However, there will also be some microscopic...

Claims

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Application Information

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IPC IPC(8): A61K39/12A61K39/39A61K9/113A61P31/14A61P37/04A61P29/00A61K31/59A61K31/355A61K31/525A61K38/06A61K36/90A61K36/896
CPCA61K39/12A61K9/0002A61K9/113A61K31/355A61K31/525A61K31/59A61K36/287A61K36/515A61K36/539A61K36/804A61K36/808A61K36/85A61K36/896A61K36/90A61K38/06A61K39/39A61K2039/5252A61K2039/552A61K2039/55566C12N2720/10034A61K2300/00
Inventor 沈建军张秀文李阳
Owner 浙江美保龙生物技术有限公司
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