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Preparation method of Brucellosis live vaccine and product thereof

A technology for brucellosis and live vaccines, which is applied in the production field of brucellosis vaccines, can solve the problems of high production cost and low yield, and achieve the effects of low production cost, high yield and increased number of cultured bacteria

Active Publication Date: 2013-03-27
哈药集团生物疫苗有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] The technical problem to be solved by this invention is to overcome the problems of low yield and high production cost in the existing production method of Brucella live vaccine, and provide a new production method of Brucella live vaccine. The number of bacteria obtained from culture can reach 150-180 billion / ml, which has the advantages of high yield and low production cost

Method used

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  • Preparation method of Brucellosis live vaccine and product thereof

Examples

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Comparison scheme
Effect test

Embodiment 1

[0021] Preparation of primary strains: After unpacking, live brucellosis vaccine A19 strain freeze-dried strains (purchased from the China Veterinary Drug Administration) were dissolved in peptone water and transplanted on tryptone agar plates or other suitable media by streaking. Cultured at 37°C for 3 days. Select more than 10 qualified colonies, transplant them on tryptone agar slant medium, cultivate them at 37°C for 72 hours, and store them as primary seeds at 2-8°C;

[0022] Preparation of secondary strains: Take primary strains and inoculate them on tryptone agar flats or other suitable media, culture them at 37°C for 72 hours, check the purity with the naked eye, add appropriate amount of peptone water to each flat, wash the bacterial lawn, and inoculate Cultivate in 10,000 ml of Martin Broth liquid culture medium at 37°C for 72 hours, obtain secondary strains after pure inspection, and store at 2-8°C; the composition of each 1000ml Martin Broth Medium is:

[0023] Be...

Embodiment 2

[0030] Preparation of primary strains: After unpacking, live brucellosis vaccine A19 strain freeze-dried strains (purchased from the China Veterinary Drug Administration) were dissolved in peptone water and transplanted on tryptone agar plates or other suitable media by streaking. Cultured at 37°C for 3 days. Select more than 10 qualified colonies, transplant them on tryptone agar slant medium, cultivate them at 37°C for 72 hours, and store them as primary seeds at 2-8°C;

[0031] Preparation of secondary strains: Take primary strains and inoculate them on tryptone agar flats or other suitable media, culture them at 37°C for 72 hours, check the purity with the naked eye, add appropriate amount of peptone water to each flat, wash the bacterial lawn, and inoculate Incubate in 10,000 ml of Martin Broth liquid medium at 37°C for 72 hours, obtain secondary strains after pure inspection, and store at 2-8°C;

[0032] Bacteria culture: add 0.01% antifoaming agent according to the qua...

Embodiment 3

[0034] Preparation of primary strains: After unpacking, live brucellosis vaccine A19 strain freeze-dried strains (purchased from the China Veterinary Drug Administration) were dissolved in peptone water and transplanted on tryptone agar plates or other suitable media by streaking. Cultured at 37°C for 3 days. Select more than 10 qualified colonies, transplant them on tryptone agar slant medium, cultivate them at 37°C for 72 hours, and store them as primary seeds at 2-8°C;

[0035]Preparation of secondary strains: Take primary strains and inoculate them on tryptone agar flats or other suitable media, culture them at 37°C for 72 hours, check the purity with the naked eye, add appropriate amount of peptone water to each flat, wash the bacterial lawn, and inoculate Cultivate in 20,000 ml of Martin Broth liquid medium at 37°C for 72 hours, obtain secondary strains after pure inspection, and store at 2-8°C;

[0036] Bacteria culture: add 0.01% antifoaming agent according to the qua...

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Abstract

The invention discloses a preparation method of a Brucellosis live vaccine and a product thereof. The method comprises primary strain preparation, secondary strain preparation and bacterial solution culture, wherein in the step of bacterial solution culture, the secondary strain is inoculated onto a liquid culture medium, and air is introduced in a manner of gradually increasing ventilation for culture; and after the bacteria strain is cultured for 30 hours, pure oxygen is introduced till the bacterial strain is cultured for 36 hours. In the method disclosed by the invention, pure oxygen is introduced at a time when it is most suitable for bacteria to prorogate in a large amount in the bacterial solution culture step, so the quantity of the cultured bacteria is increased greatly to 150 to190 billion per milliliter. When the method is used, the bacterial solution is not required to be concentrated; and the method has the advantages of high yield, low production cost and the like.

Description

technical field [0001] The invention relates to a preparation method of an animal vaccine, in particular to a preparation method of a brucellosis vaccine and a vaccine prepared by the method, and belongs to the production field of the brucellosis vaccine. Background technique [0002] Brucellosis (full name brucellosis) is a zoonotic acute and chronic infectious disease caused by Brucella. The clinical features are slow onset, long-term fever, sweating, weakness, general pain and joint pain , Symptoms in the acute phase usually subside within 3 to 6 months. Brucellosis is divided into six types: cattle, sheep, pigs, squirrels, sheep and canine Brucella. The first three are mainly found in China. [0003] Brucella is a small, short rod-shaped or club-shaped, non-spore-forming, Gram-negative bacillus. Brucella is sensitive to heat and can die at 70°C for 10 minutes; it will die under direct sunlight for 1 hour; it loses its vitality rapidly in spoilage materials; commonly use...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/10C12N1/20A61P31/04
Inventor 张继东付丽杰尹尧郑铁鑫王华柴华袁明霞
Owner 哈药集团生物疫苗有限公司
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