Preparation technology of mycoplasma ovipneumoniae inactivated vaccine

A Mycoplasma pneumoniae, preparation technology, applied in the direction of medical preparations containing active ingredients, bacterial antigen components, antibody medical components, etc.

Inactive Publication Date: 2013-05-22
SOUTHWEST UNIVERSITY FOR NATIONALITIES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are no reports on the preparation of inactivated Mycoplasma pneumoniae vaccine to remove antigen endotoxin and evaluate the efficacy of different adjuvants in immune adjuvant

Method used

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  • Preparation technology of mycoplasma ovipneumoniae inactivated vaccine
  • Preparation technology of mycoplasma ovipneumoniae inactivated vaccine

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0012] (1) Preparation of medium

[0013] Modified Thiaucourt's medium was used. The medium components are as follows: 21.0 g of PPLO broth, 100 mL of fresh yeast extract, 200 mL of inactivated horse serum, 2 mL of 0.5 g / ml glucose, 8 mL of 0.5 g / mL sodium pyruvate, 3.2 mL of 0.5% phenol red, 10 ml of 10000U / mL penicillin, 0.1 g of thallium acetate, make up to 1000 ml with deionized water, adjust the pH to 7.6-7.8, filter and sterilize with a 0.22 μm bacterial filter, aliquot and store at 4°C for later use.

[0014] (2) Proliferation of Mycoplasma pneumoniae

[0015] Add 1 mL of modified Thiaucourt's medium to dissolve the lyophilized bacteria of Mycoplasma ovis pneumoniae, add 1 mL of bacterial liquid to 10 mL of modified Thiaucourt's medium, and culture on a shaker at 37 C°160 rpm for 48 h (the color of the medium gradually changes from red to (Orange yellow, clear and translucent bacterial liquid, sterile film, no turbidity); inoculate the first-generation culture into...

Embodiment 1

[0031] Example 1 Determination of Antigen Endotoxin, Vaccine Safety Test and Effectiveness Test

[0032] (1) Determination of endotoxin

[0033] The content of endotoxin in inactivated antigen of Mycoplasma ovis pneumoniae was determined by dynamic turbidity method, and the results showed that the content of endotoxin in antigen decreased greatly after potassium dichromate treatment. The endotoxin content of the antigen not treated with potassium dichromate was 912 EU / mL, and the endotoxin content of the antigen treated with potassium dichromate was only 38 EU / mL, that is to say, after being treated with potassium dichromate, the antigen endotoxin content The toxin content was reduced by 24 times, improving the safety of the vaccine.

[0034] (2) Safety inspection

[0035] 5 healthy Chengdu Ma sheep around 5 months old (use MO indirect hemagglutination diagnostic kit to detect no antibodies, use nasal swabs to extract DNA, and PCR identified no MO), neck muscle injection o...

Embodiment 2

[0038] Example 2 Screening of vaccine adjuvants

[0039] Select adjuvants commonly used in inactivated vaccines: white oil adjuvant, ISA-206 adjuvant, Buonavoglial's adjuvant, and ISA-760 adjuvant to prepare vaccines with inactivated antigens respectively, and immunize healthy adult rabbits by detecting their serum antibody levels , to evaluate the adjuvant effect of the adjuvant on the vaccine, and to screen out the ideal inactivated vaccine adjuvant.

[0040] method:

[0041] 1. Antigen preparation: Antigen preparation was carried out according to the method described in the specific embodiment.

[0042] 2. Preparation of inactivated vaccine:

[0043] (1) Preparation of inactivated vaccine with white oil adjuvant: Add sterilized Tween-80 with a final concentration of 3% according to the volume of antigen, stir while adding, until completely dissolved, and make the aqueous phase of inactivated vaccine. The volume ratio of water phase and white oil adjuvant is 1:1. ...

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Abstract

The invention discloses a preparation technology of a mycoplasma ovipneumoniae inactivated vaccine, which is based on the improvement on the preparation technology of the mycoplasma ovipneumoniae inactivated vaccine, and provides a preparation method of the mycoplasma ovipneumoniae inactivated vaccine. Through the application of the method provided by the invention, an ideal efficient mycoplasma ovipneumoniae inactivated vaccine adjuvant is screened, the content of toxin in antigen is greatly reduced, and the vaccine safety is improved. After the vaccine is applied to sheep, the antibody is generated early, the titer is high, the duration is long, and the mycoplasma ovipneumoniae infection can be effectively prevented and controlled.

Description

technical field [0001] The invention relates to an inactivated vaccine of Mycoplasma pneumoniae in veterinary biological products, in particular to a preparation process of Mycoplasma pneumoniae inactivated vaccine. Background technique [0002] Mycoplasma ovipneumoniae (MO) causes atypical pneumonia in sheep and goats, clinically characterized by runny nose, cough, progressive emaciation and proliferative interstitial pneumonia. In 1963, Mackay et al. isolated MO for the first time from the lungs of sheep with pneumonia. Since then, MO infection has been reported to cause disease in sheep and goats in many other countries and regions. At present, Mycoplasma ovis pneumoniae is distributed all over the world. In recent years, the incidence of the disease has been increasing year by year, causing huge economic losses to the sheep industry. In addition, the susceptibility of sheep to other pathogens (such as Pasteurella multocida, Mannella hemolyticus and influenza virus) w...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/107A61K39/39A61K39/02A61P31/04C12N1/36
Inventor 汤承岳华费磊
Owner SOUTHWEST UNIVERSITY FOR NATIONALITIES
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