Preparation method of serum-free mycoplasma ovipneumoniae antigen

A Mycoplasma pneumoniae, serum-free technology, applied in the directions of microorganism-based methods, biochemical equipment and methods, bacterial antigen components, etc., can solve the problem of lack of Mycoplasma sheep pneumoniae antigen, etc., and achieve the effect of convenient and simple operation.

Pending Publication Date: 2019-11-15
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Preparation method of serum-free mycoplasma ovipneumoniae antigen
  • Preparation method of serum-free mycoplasma ovipneumoniae antigen

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Embodiment 1

[0029] A preparation method of serum-free Mycoplasma pneumoniae antigen is as follows:

[0030] (1) Chicken embryo preparation: Prepare 7-day-old healthy SPF chicken embryos (SPF eggs were purchased from Shandong Haotai Experimental Animal Breeding Co., Ltd.). 50%).

[0031] (2) Preparation of Mycoplasma pneumoniae inoculum: resuscitate Mycoplasma pneumoniae Y98 strain (purchased from China Veterinary Microorganism Culture Collection Center CVCC, No. CVCC384), culture in a 37°C incubator for 1 day, subculture, centrifuge at 8000rpm for 15min, and centrifuge in normal saline After washing the cultured bacteria solution, resuspend it with an equal volume of normal saline to obtain a Mycoplasma ovis pneumoniae inoculum.

[0032] (3) Chicken embryo inoculation, cultivation and harvest: inoculate 7-day-old healthy SPF chicken embryos with the Mycoplasma ovis pneumonia inoculum, inoculate through the yolk sac, and inoculate 0.2ml of each chicken embryo. After the inoculation, the i...

Embodiment 2

[0035] A preparation method of serum-free Mycoplasma pneumoniae antigen is as follows:

[0036] (1) Chicken embryo preparation: Prepare 7-day-old healthy SPF chicken embryos (SPF eggs were purchased from Shandong Haotai Experimental Animal Breeding Co., Ltd.). 50%).

[0037] (2) Preparation of Mycoplasma pneumoniae inoculum: resuscitate Mycoplasma pneumoniae Y98 strain (purchased from China Veterinary Microorganism Culture Collection Center CVCC, No. CVCC384), culture in a 37°C incubator for 1 day, subculture, centrifuge at 8000rpm for 15min, and centrifuge in normal saline After washing the culture solution, resuspend it with an equal volume of normal saline to obtain the inoculum.

[0038] (3) Chicken embryo inoculation, cultivation and harvest: 7-day-old healthy SPF chicken embryos were inoculated with the Mycoplasma ovis pneumonia inoculum, inoculated through the yolk sac, and each chicken embryo was inoculated with 0.2ml. After the inoculation, the incubator continued to...

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Abstract

The invention provides a preparation method of serum-free mycoplasma ovipneumoniae antigen. The method comprises the following steps: (1) preparation of chicken embryos: preparing the healthy SPF chicken embryos at 7-8 days of age, the embryos are vigorous, clear in blood vessels, and cultured in an incubator; (2) preparation of mycoplasma ovipneumoniae inoculum; and (3) chicken embryo inoculation, culture and harvest: inoculating the healthy SPF chicken embryos of 7-8 days old in step (1) of the mycoplasma ovipneumoniae inoculum, 0.2ml of the inoculum is inoculated for each chicken embryo, After inoculation, the incubator is further cultured for 5 days, and the allantoic fluid of dead chicken embryos and/or the allantoic fluid of chicken embryos that do not died after 5 days are aseptically obtained to obtain the serum-free mycoplasma ovipneumoniae antigen. In the present invention, the mycoplasma ovipneumoniae is subcultured in chicken embryos, and the allantoic fluid of chicken embryos is collected to obtain the serum-free mycoplasma ovipneumoniae antigen. The method of the present invention does not use serum, and can avoid many problems in using serum.

Description

technical field [0001] The invention belongs to the technical field of veterinary biotechnology, and in particular relates to a preparation method of serum-free sheep mycoplasma pneumoniae antigen. Background technique [0002] Mycoplasma ovis pneumoniae can cause pneumonia in sheep and goats. Clinically, the main symptoms are runny nose and cough. Mycoplasma ovis pneumoniae exists widely in sheep, and the pathogen can also be isolated from the upper respiratory tract of normal sheep. Symptoms tend to be exacerbated by stressful stimuli such as weather changes, transportation, or secondary to other bacteria or viruses. In recent years, the number of diseases caused by or secondary to Mycoplasma pneumoniae in sheep farms in my country has increased, seriously threatening the healthy development of the sheep industry. [0003] Currently, modified KM2 medium and modified Thiaucourt's medium are mainly used for culture of Mycoplasma pneumoniae antigen in sheep. In the existin...

Claims

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Application Information

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IPC IPC(8): C12N1/20A61K39/02A61P31/04C12R1/35
CPCA61K39/0241A61P31/04C12N1/20
Inventor 郝华芳储岳峰陈胜利颜新敏
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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