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Method for preparing serum-free mycoplasma bovis antigen

A Mycoplasma bovis, serum-free technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, bacterial antigen components, etc., can solve the problems of pollution, bovine body affecting the immune effect, serum increasing the cost of vaccine production, etc., to achieve stable sources , easy to operate, easy to obtain the effect

Pending Publication Date: 2019-12-10
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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Problems solved by technology

The complexity of serum components and the differences between batches affect the quality control of vaccines. Serum is easily contaminated by mycoplasma and other microorganisms. In the later stage, a purification process is required to remove complex components of serum. Serum antigens are likely to increase the side effects on cattle and affect the immune effect. In addition, the use of serum Increased seedling cost

Method used

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  • Method for preparing serum-free mycoplasma bovis antigen
  • Method for preparing serum-free mycoplasma bovis antigen

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Embodiment 1

[0028] A method for preparing Mycoplasma bovis antigen without serum is as follows:

[0029] (1) Put fresh SPF eggs in an incubator to incubate (temperature 37.0-37.5°C, humidity 50%), cultivate for 7 days, illuminate the eggs before inoculation, pick healthy chicken embryos with well-developed, clear blood vessels, and vigorous vitality, discard Remove non-fertilized eggs, dead embryos and weak embryos, draw the air cell position of healthy chicken embryos in the dark room, and reserve the chicken embryos.

[0030] (2) Preparation of Mycoplasma bovis inoculum: resuscitate and subculture Mycoplasma bovis PG45 strain (ATCC, No. ATCC 25523), culture in a constant temperature incubator at 37°C for 24 hours, the medium is modified Thiaucourt's medium, and the color of the medium is determined. Turn yellow (or when the pH value drops from 7.4 to 7.0), centrifuge the fresh Mycoplasma bovis culture bacterium liquid, centrifuge 15min at 8000rpm, resuspend the thalline with equal volum...

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Abstract

The invention provides a method for preparing a serum-free mycoplasma bovis antigen. The method for preparing the serum-free mycoplasma bovis antigen comprises the following steps: (1) hatching freshSPF breeding eggs, carrying out culturing for 7-8 days, and selecting healthy SPF chicken embryos; (2) preparing a mycoplasma bovis inoculum; (3) inoculating the selected healthy SPF chicken embryos,which are 7-8 days old, with the mycoplasma bovis inoculum, continuing culturing after the inoculation in an incubator, observing status of the SPF chicken embryos, collecting dead inoculated SPF chicken embryos, and carrying out storing at 2-8 DEG C for 12-24 hours; and (4) aseptically collecting allantoic fluid of the dead SPF chicken embryos, thereby obtaining the serum-free mycoplasma bovis antigen. The method for preparing the serum-free mycoplasma bovis antigen is capable of realizing serum-free production of mycoplasma bovis antige, thereby avoiding multiple problems caused by use of animal serum; and moreover, the cultured mycoplasma bovis antigen is high in live bacterium titer.

Description

technical field [0001] The invention relates to a serum-free method for preparing mycoplasma bovis antigen. Background technique [0002] Mycoplasma bovis is an animal disease that seriously endangers the cattle industry in the world. The disease is caused by Mycoplasma bovis (M.bovis), and the main clinical manifestations are calf pneumonia, arthritis, and mastitis in dairy cows. The disease is currently endemic worldwide causing significant economic losses. Antibiotic therapy and vaccine immunization are important means to prevent and control bovine mycoplasma disease. In recent years, the emergence of more and more drug-resistant strains has brought serious challenges to the prevention and control of bovine mycoplasma disease. [0003] Antigen production is a key link in vaccine preparation and affects vaccine quality. At present, Mycoplasma bovis antigen production and cultivation mainly use Mycoplasma bovis liquid medium, such as improved KM2 medium, Niuxintang medi...

Claims

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Application Information

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IPC IPC(8): C12N1/20A61K39/02A61P31/04C12R1/35
CPCA61K39/0241A61K2039/552A61P31/04C12N1/20
Inventor 陈胜利储岳峰郝华芳颜新敏刘永生
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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