Application of Mycoplasma capricolum subsp. capricepneumonia mccp1801 in preparation of goat infectious pleuropneumoniae vaccine
A technology for pleuropneumonia and mycoplasma, applied in the direction of antibody medical components, bacterial antigen components, medical preparations containing active ingredients, etc., can solve problems such as limited protective effect, inactivation of antigenic epitopes, effective antigen destruction, etc., and achieve growth performance Excellent, early antibody production, improved immune response
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[0027] The method for preparing the inactivated antigen of the present invention preferably comprises: (1) inoculating the recovered Mycoplasma capricoides subsp. The improved MTB culture medium includes components at 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 water respecti...
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]8-9 month old healthy goats (Mccp antibody and antigen negative) were inoculated with the inactivated vaccine of Mycoplasma capricolum subspecies. Each goat was injected with 2 ml of the vaccine in the neck muscles, and the control group was injected with the same amount of PBS for 14 days of continuous observation. Results The goats in the vaccine group and the control group were both healthy and alive, the sheep had no symptoms such as coughing and sneezing, no abnormality in feeding and drinking, and no obvious lumps and other adverse reactions in the injected part, and the vaccine of the present invention was safe.
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