细胞内快速连续传代培养甲肝病毒的研究  

STUDY ON RAPID AND CONSECUTIVE CULTURE OF HAV IN KMB17 CELLS

作　　者：谢忠平[1] 宋霞[1] 全文琦[1] 熊秋霞[1] 段维国[1] 丁雪凤[1] 王燕[1] 邹静如[1] 

机构地区：[1]中国医学科学院中国协和医科大学医学生物学研究所,昆明650118

出　　处：《现代预防医学》2006年第7期1089-1090,共2页Modern Preventive Medicine

摘　　要：目的：研究甲肝病毒在细胞内快速连续传代培养病毒增殖情况，探索疫苗生产工艺改进的可能性。方法：将甲肝病毒吕8株感染KMB17细胞后形成带毒细胞，每7d传代1次，检测每次收获物的甲肝病毒抗原（HAAg）滴度及感染性滴度，比较滴度的变化，同时检测一步生长曲线，分析病毒增殖特性有无变化。结果：甲肝病毒吕8株感染KMB17细胞后每7d传代1次，其第1～8代抗原滴度为1024-2048Eu／0．1ml，感染性滴度为7．33～7．67lgCCID50／ml，第9代开始下降，抗原滴度仅为256Eu／0．1ml，感染性滴度为6．83～7．00 lgCCID50／ml；在连续传10代内，细胞均未出现CPE现象；第10代与第1代相比，其一步生长曲线病毒增殖高峰均为20-25d，没有明显变化。结论：甲肝病毒吕8株在KMB17细胞上每7d传代1次，在第8代以内，其抗原滴度及感染性滴度均能达到较高的程度。经细胞内连续快速传代后．HAV吕8株生长的增殖高峰约为第20-25d，与传代前样品没有明显差别。Objective： To study the proliferation of HAV which were cultured rapidly and consecutively in cells, and to investigate the possibility of improving the production process of inactivated HAV vaccine. Methods： The KMB17 cells were infected by HAV L8 strain and the cell passage was performed once every 7 days. The HAAg content and infectious titers were tested in each harvest and the titer variation was compared. Meanwhile, the one-step growth curve was tested to analyze whether the virus proliferation changed or not. Results： The HAAg content was 1 024-2 048 Eu/0.1 ml and the infection titer was 7.33-7.67 lg CCID50/ml from passage 1 to passage 8, but the injection titer and the HAAg content began to decrease from passage 9 with the HAAg content of 256 Eu/0.1 ml and the infectious titer of 6.83-7.00 lg CCID50/ml, respectively. The CPE was not noticed during the consecutive 10 passages. Compared passage 10with passage 1, the virus proliferation peak of one - step growth curve is 20 to 25 days without obvious variation. Conclusion： Within the passage 8 of HAV L8 strain in KMB17 cells, the HAAg content and infectious titer could reach to a higher level, which can be used for vaccine production process. After rapid and consecutive passages in KMB17 cells, the proliferation peak of HAV L8 strain is about 20 to 25 days, which makes no significant differences with the samples before passage.

关 键 词：甲型肝炎病毒 细胞内培养 生长曲线 

分 类 号：R373.21[医药卫生—病原生物学]