重组禽流感病毒H5亚型Re-7株在MDCK细胞上增殖条件的研究

摘要

In order to explore the value increment law of the H5 subtype Re-7 strains in MDCK cells,the optimal dose and the optimal harvest time were determined. To restructure the H5 virus subtype Re-7 strains inoculation to 100 L of bioreactor full suspension MDCK cells proliferation of serum-free culture experiment, testing several different virus infection, virus HA, TCID50and different time after inoculation EID50. The virus was inoculated into MDCK cells with the best value added conditions.Determine the optimum dose MOI for 10-2, and the best harvest time is 60 h. In the 100 L bioreactor,repeated verification was conducted to obtain stable test results. The virus HA reached 1 : 1024, and the content of every 1mL virus reached 107.33TCID50, and the content of every 0.1 mL virus reached 106.83EID50.This study provides a relatively stable parameter index for the recombinant avian influenza virus H5 subtype Re-7.%为了探索重组禽流感病毒H5亚型Re-7株在MDCK细胞上的增值规律,确定最适的接毒量与最佳收获时间.将重组禽流感病毒H5亚型Re-7株接种至100 L生物反应器全悬浮无血清培养的MDCK细胞进行增殖试验,检测不同病毒感染量,接种后不同时间病毒的HA、TCID50以及EID50.根据确定的最佳增值条件将病毒接种到MDCK细胞中进行大规模增殖培养.确定最适接毒量MOI为10-2,最佳收获时间为60 h.在100 L生物反应器中进行重复验证,获得稳定的试验结果,病毒HA达到1 : 1024,每1 mL病毒含量达到107.33TCID50,每0.1 mL病毒含量达到106.83EID50.研究为重组禽流感病毒H5亚型Re-7株的全悬浮规模化生产提供了相对稳定的参数指标.