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Application of mycoplasma capricolum subsp. Capripneumoniae Mccp1801 in preparation of contagious caprine pleuropneumonia vaccine

A pleuropneumonia and mycoplasma technology, applied in bacterial antigen components, antibody medical components, medical preparations containing active ingredients, etc., can solve problems such as limited protection effect, inactivation of antigenic epitopes, effective antigen destruction, etc., and achieve growth performance. Excellent, early antibody production time, the effect of improving immune response

Active Publication Date: 2021-06-04
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the bacteria inactivated vaccines are made from strains that were popular more than ten years ago or earlier. They are inactivated by formaldehyde and other inactivators, and are made with adjuvants such as oil adjuvants. Some sheep have relatively large side effects after immunization. The problem of effective antigen destruction or antigenic epitope inactivation in inactivators such as formaldehyde, as well as the limited protective effect and high cost of virulent Mycoplasma capricosum goat pneumonia subspecies infection

Method used

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  • Application of mycoplasma capricolum subsp. Capripneumoniae Mccp1801 in preparation of contagious caprine pleuropneumonia vaccine
  • Application of mycoplasma capricolum subsp. Capripneumoniae Mccp1801 in preparation of contagious caprine pleuropneumonia vaccine
  • Application of mycoplasma capricolum subsp. Capripneumoniae Mccp1801 in preparation of contagious caprine pleuropneumonia vaccine

Examples

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preparation example Construction

[0027] The preparation method of the inactivated antigen of the present invention preferably comprises: (1) inoculating the bacterial strain of Mycoplasma capricosum subsp. The improved MTB culture medium includes components of the following concentrations: 2 g / L of glucose, 2 g / L of sodium pyruvate, 21 g / L of PPLO broth, 100 ml / L of yeast extract with a mass percentage of 25%, and a mass percentage of 1% phenol red 2.5ml / L, horse serum 200ml / L, 100,000 IU penicillin, pH 7.4-7.6;

[0028] (2) centrifuging the mycoplasma bacterium liquid, collecting the bacterium precipitate, and resuspending to obtain a mycoplasma antigen solution with a concentration of 100-400 μg / ml;

[0029] (3) Mix the mycoplasma antigen solution with saponin and inactivate at 4°C for 24-48 hours to obtain the inactivated antigen.

[0030]When the present invention prepares the improved MTB medium described in step (1), it is preferred to dissolve 2 g of glucose and 2 g of sodium pyruvate in 25 ml of wate...

Embodiment 1

[0042] Inactivated vaccine preparation

[0043] (1) Preparation of antigen solution

[0044] 1) Medium preparation method: Dissolve 2g of glucose and 2g of sodium pyruvate in 25ml of water respectively, filter and sterilize with a 0.22μm microporous membrane for use; dissolve 21g of PPLO broth in 650ml of deionized water, add 100ml of 25% yeast extract , 2.5ml of 1% phenol red, mixed evenly, sterilized by autoclaving at 121°C for 20min, after cooling, add 200ml of sterile healthy horse serum, 25ml of 8% glucose and 25ml of 8% sodium pyruvate solution sterilized by filtration and 100,000 IU / ml penicillin 1ml, adjust the pH value to 7.4-7.6 with sterilized 1mol / L NaOH, aseptically dispense, store at 4°C for later use, that is, the improved MTB medium.

[0045] 2) Mycoplasma antigen preparation:

[0046] Resuscitate Mycoplasma goati subspecies M1801, inoculate into 4.5ml of improved MTB medium, and culture at 37°C for 24 hours. When the pH value of the culture drops to 6.5-6.8,...

Embodiment 2

[0057] Safety Test of Inactivated Vaccine

[0058]Healthy goats aged 8 to 9 months (Mccp antibody and antigen negative) were vaccinated with the inactivated vaccine of Mycoplasma capricum subspecies goat pneumonia. Each goat was injected with 2ml of the vaccine in the neck muscle, and the control group was injected with the same amount of PBS, and the observation continued for 14 days. Results The goats in the vaccine group and the control group were healthy and alive. The goats had no symptoms such as coughing and sneezing, no abnormality in eating and drinking, and no adverse reactions such as obvious lumps in the injected part. The vaccine of the present invention was safe.

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Abstract

The invention provides application of mycoplasma capricolum subsp. Capripneumoniae Mccp 1801 in preparation of a contagious caprine pleuropneumonia vaccine, and relates to the technical field of biology. The invention further provides a vaccine based on the M1801 strain and a preparation method thereof. The vaccine can improve the immune protection effect, reduce immune side reactions and provide a good immune protection effect; meanwhile, the M1801 strain is high in virulence, good in immunogenicity and excellent in growth performance, a body can be induced to generate a high-level antibody when the strain is used for preparing a vaccine, and the antibody is early in generation time and high in titer; the vaccine safety is high; and immune sheep is protected by 4 / 5 or more, and symptoms such as body temperature rise, cough, sneezing, depressed spirit or tissue lesions can be obviously relieved.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to the application of Mycoplasma capricosum capres pneumoniae subspecies Mccp 1801 in the preparation of goat infectious pleuropneumonia vaccine. Background technique [0002] Contagious caprine pleuropneumonia (CCPP) is a contagious respiratory infectious disease of goats, caused by Mycoplasma capricolum subsp. Fibrinous pleuropneumonia and hepatic changes in the lungs are one of the infectious diseases notifiable by the World Organization for Animal Health (OIE). Goat infectious pleuropneumonia is mainly transmitted by contact and droplets. When the disease breaks out in a new epidemic area, the incidence rate is as high as 100%, and the mortality rate can be as high as 80%. Africa, the Middle East, Asia and other regions are the main epidemic areas of the disease. In recent years, the range of hosts infected by the disease has been increasing, and it has become a serious...

Claims

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Application Information

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IPC IPC(8): A61K39/02A61K39/39A61P31/04
CPCA61K39/0241A61K39/39A61K2039/521A61K2039/55577A61K2039/575A61P31/04
Inventor 储岳峰郝华芳陈胜利颜新敏马丽娜刘永生
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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