仙台病毒高滴度病毒制备方法的建立

目的：建立使用传代细胞稳定大量地制备高滴度的仙台病毒（SeV）的方法，取代成本高、周期长、操作繁琐、易污染的鸡胚制备仙台病毒的方法，用于实验研究。方法：首先观测不同细胞感染仙台病毒后发生病变的时间、程度以及产毒量，确定仙台病毒敏感细胞株并确定病毒产量高峰期，连续在细胞中传3代测量上述指标。其次观测染毒时病毒吸附时间不同对病毒滴度的影响。综合分析后建立制备高滴度仙台病毒的方法。结果：大鼠脑胶质瘤细胞C6株对仙台病毒高度敏感，细胞出现病变所需时间短且明显，病毒产量最高。仙台病毒滴度与染毒时的吸附时间呈正相关，病毒最佳吸附时间为90min。接种病毒后72h收获的病毒滴度最高，平均为11 TCID50，并且病毒可以在细胞内连续稳定传代。结论：确认大鼠脑胶质瘤细胞C6株为仙台病毒的敏感细胞。且病变快、明显，病毒产量高。为在体外深入研究仙台病毒和大量繁殖高滴度仙台病毒提供了实验方法。

A preparative method for producing high titer Sendai virns

Objective： To establish a preparative method of producing high titer Sendai virus using passage cells in empirical study so that it can supersede the one of inoculating chick embryo eggs. Methods： First of all, score of cytopatho effect （CPE） and yield were observed after three cell lines to be infected with Sendai virus. And then the cell lines sensitive to Sendal virus and the peak yield of resulting virus were determined. These data were obtained in every passage throughout three consecutive passages. Secondly, the impact of different viral adsorption time on the yield was investigated. Thirdly, a preparative method of producing high titer Sendai virus was established by comprehensive analysis. Results：The variables mentioned above showed that rat neurogliocytoma C6 was hypersensitive to Sendai virus with apparent CPE and higher yield in different cells tested. The virus titer was adsorption-time dependent. Furthermore, the optimum viral adsorption time was about 90 minutes, and the peak titer appears at 72 hours after viral adsorption and the average titer was 11 T CID50 . Moreover, Sendai virus could be passaged in rat neuroglioeytoma C6 stably. Conclusion： Rat neurogliocytoma C6 is confirmed to be the sensitive cell lines to Sendai virus because of apparent CPE, high virus yield. Thus it is demonstrated that the method established will paves the way for producing high titer Sendai virus and therefore elaborating the virus in vitro in the future.