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The Research of Cold Adaptation of Influenza Virus H3N2 in Vero Cell

Author: DingShiLei
Tutor: LiaoGuoYang；JiangShuDe；LiWeiDong；SunMingBo
School: Peking Union Medical College , China
Course: Pathogen Biology
Keywords: Cold-adapted influenza vaccine Vero cells M-gene
CLC: S852.65
Type: Master's thesis
Year: 2008
Downloads: 73
Quote: 0
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Abstract


Influenza is a respiratory infection caused by influenza virus (influenza virus) , infection , and a variety of animals , which is characterized by the sudden outbreak of the short term , quickly became popular , with a wide range of destructive . Prevention and treatment of influenza is influenza vaccination . The influenza vaccine market in the chick embryo production matrix inactivated vaccine , limited to the limitations of the chick embryo production of influenza vaccine , developed cells influenza vaccine to become a top priority . The Netherlands has ratified the MDCK cell -based influenza vaccine production , but Vero cells the WHO actively recommended for vaccine production cell matrix . Cold-adapted live influenza vaccine , live attenuated vaccine because of its superiority and attention . The study based on the adaptation of influenza virus strains in Vero cells at 25 ℃ continuous passage in order to obtain the cold-adapted virus strains , and cells for the production of cold-adapted influenza vaccine related preparations . The first part of the flu virus will adapt to the Explore this experiment in Vero cells on 25 ° C cold adapted Vero cells with the influenza virus strains cultured at 25 ℃ observed virus culture fluid in the different concentration of TPCK- trypsin , NaHCO 3 the impact of concentration on virus proliferation , screening out the 25 ℃ viruses suitable culture conditions . Culture conditions , blind passages , temperature 33 ° C normal cultured Reply culture virus mixed cultivation of a variety of methods , 25 ° C subculture . Passage virus hemagglutinin titer by Generations reduced undetectable hemagglutination titer to the 7th generation by direct cold - adapted strains of the virus failed to obtain the virus from 33 ℃ to 25 ℃ adapt . The gradient of the second part of the influenza viruses on Vero cells cold adapted virus seed of the 33 ° C harvested at 28 ℃ first to adapt, to be stable titer and continued to obtain a relatively high titer of the virus after 28 ° C for adaptation as a toxic species . Transferred to 25 ° C for adaptation , this temperature gradient to adapt to the strains able to increase growth in 25 ℃ passages to get a low-level cold-adapted strain of the virus in 25 ℃ continuous passage for more than 15 generations . Reply 33 ℃ culture incubated at 25 ℃ for more than 15 generations still high titer , indicating that the strain has not temperature sensitive characteristics , needs further subcultured at 25 ℃ . In this test , using the method of detecting virus M gene to determine the presence of the virus .

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CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Animal Medicine ( Veterinary Medicine) > Basic Veterinary Science > Animal Microbiology ( Veterinary Microbiology, ) > Livestock Virology
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