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A technology of attenuated live vaccine, varicella, applied in the field of medical microbiology
Active Publication Date: 2008-04-16
SHANGHAI RONGSHENG BIOLOGICAL PHARM CO LTD
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Problems solved by technology
Although the chickenpox vaccine market in China is still in its infancy, the market potential is huge
Method used
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Embodiment 1
[0040] A live attenuated varicella vaccine, prepared as follows:
[0041] (1) MRC-5 cells were continuously passaged for 20 generations in cell flasks;
[0042] (2) After the cells grow to a certain scale, the virus seed is inoculated into the cells, and two consecutive infections are carried out, and the M.O.I is respectively 0.001 and 0.01 to expand the harvest of the virus liquid;
[0043] (3) adding MEM culture solution containing 2% fetal bovine serum, and culturing at 35°C;
[0044] (4) When the lesion degree of MRC-5 cells reaches ++~+++, pour out the culture medium, wash the cell surface with PBS washing solution, and wash away the bovine serum;
[0045] (5) Add 0.05% EDTA solution to elute the infected cells, collect the cell suspension in a centrifuge tube, centrifuge at 1800 rpm for 5 minutes at 4°C, discard the supernatant, add the vaccine solution containing a stabilizer, mix well, and collect , aspirated for single-bottle sterility testing, and stored at -80°C;...
Embodiment 2
[0052] A live attenuated varicella vaccine, prepared as follows:
[0053] (1) MRC-5 cells are continuously passaged in cell flasks for no more than 25 generations;
[0054] (2) After the cells grow to a certain scale, the virus seed is inoculated into the cells, and two consecutive infections are carried out, and the M.O.I is respectively 0.0015 and 0.02, so as to expand the harvest of the virus liquid;
[0055] (3) Add MEM culture solution containing 2.5% fetal bovine serum, and culture at 36°C;
[0056] (4) When the lesion degree of MRC-5 cells reaches ++~+++, pour out the culture medium, wash the cell surface with PBS washing solution, and wash away the bovine serum;
[0057] (5) Add 0.05% EDTA solution to elute the infected cells, collect the cell suspension in a centrifuge tube, centrifuge at 1800 rpm for 5 minutes at 6°C, discard the supernatant, add the vaccine solution containing a stabilizer, mix well, and collect , aspirated for single-bottle sterility testing, and...
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Abstract
The present invention relates to herpes virus family medical microorganism. The present invention is prepared by the following method: (1) serial passage MRC-5 cell; (2) inoculating to cell and continuous infecting; (3) adding culture solution; (4) washing cell surface; (5) eluting infected cell, centrifugal separating, adding vaccine fluid containing stabilizing agent to refrigerate; (6) ultrasonic crashing and centrifugal separating to acquire virus stock solution; (7) combining and diluting, and at once processing to pack and freeze-dry, preparing attenuated pox vaccine. The stabilizing agent is prepared by dissolving NaCl, KCl, KH2PO4, Na2HPO4.12H2O, saccharose, sodium glutamate, kalamycin, erythromycin and deionized water into the water with weight ratio 3:0.1:0.2:3.3:50:0.4:0.1:0.03:1000. The present invention has good advantages of safety and immunogenicity.
Description
technical field [0001] The invention relates to a medical microorganism of the herpesviridae family, in particular to a live attenuated varicella vaccine. Background technique [0002] Varicella varicella is a common respiratory infectious disease with high contact transmission caused by varicella-zoster virus (VZV), a human alpha-herpes virus that can cause two human diseases: the first infection occurs in children under 10 years old In children, causing childhood varicella, after which the virus becomes latent and often reactivates (often decades later) to produce shingles. Primary infection can cause different degrees of typical diseases in patients, especially for immunocompromised patients and those exposed to immunosuppressant therapy, the consequences are more serious. In chickenpox, the virus penetrates the peripheral nervous system and remains dormant until reactivated in the painful shingles form. Since the virus is latent, the expression of most viral genes is r...
Claims
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Application Information
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