Hircine primary atypical pneumonia combined inactivated vaccine and preparation method thereof
A dual inactivated vaccine and mycoplasma pneumonia technology, applied in the direction of bacterial antigen components, respiratory diseases, bacteria, etc., can solve the problems of indistinguishable diseases, lack of cross protection, mixed infection, etc.
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Embodiment 1
[0030] 1 Prepare culture medium
[0031] From March 11th to 12th, 2005, 42000ml of improved KM2 medium was prepared. 14700ml of 1.7% hydrolyzed milk protein Hank's buffer, 840ml of 25% yeast infusion, 420ml of 1% thallium acetate solution and 420ml of 0.4% phenol red, mixed and sterilized by 10 pounds for 20 minutes, after cooling, add 0.22μm microporous The minimum element medium MEM2 1000ml, healthy horse serum 4200ml, and 20,000 IU penicillin 420ml were sterilized by membrane filtration, the pH value was adjusted to 7.8 with 1mol / L sodium hydroxide solution, and stored at 2°C for later use.
[0032] 2 Mycoplasma proliferation
[0033] 2.1 Proliferation of Mycoplasma: On March 12, 2005, Mycoplasma goat subspecies M87-1 was inoculated with 5ml of improved KM2 medium. After 3 days of cultivation, the pH value of the culture dropped to 7.0. 5% for 3 consecutive expansion cultures, when the pH value of the culture dropped to 7.0, harvest 18000ml of mycoplasma goat subsp. 7 cc...
Embodiment 2
[0050] 1 Prepare culture medium
[0051] From April 1st to 2nd, 2005, 42000ml of improved KM2 medium was prepared. 14700ml of 1.7% hydrolyzed milk protein Hank's buffer, 840ml of 25% yeast infusion, 420ml of 1% thallium acetate solution, 420ml of 0.4% phenol red, mixed and sterilized by 10 pounds for 20 minutes, cooled and then added with a 0.22μm microporous Membrane filtration sterilized minimum element medium (MEM) 21000ml, healthy horse serum 4200ml, 20,000 IU penicillin 420ml, adjusted the pH value to 7.4 with 1mol / L sodium hydroxide solution, and stored at 8°C for later use.
[0052] 2 Mycoplasma proliferation
[0053] 2.1 Proliferation of Mycoplasma: On April 10, 2005, Mycoplasma goat subspecies M87-1 was inoculated with 10 ml of the improved KM2 medium, and after 5 days of cultivation, the pH value of the culture dropped to 6.8. Carry out 3 times of continuous expansion cultures by 5% of the total amount of the culture medium, when the pH value of the culture drops t...
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