H1N1甲型流感病毒在BHK21中的增殖规律

Study on the proliferation profile of influenza A virus in BHK21 cells

BHK21细胞是多种动物病毒的易感宿主,是病毒疫苗生产的常用基质,为了解H1N1甲型流感病毒在BHK21细胞中的增殖规律,本研究建立了定量检测甲型H1N1流感病毒M基因表达水平的SYBR GreenⅠ实时荧光RT-PCR(RRT-PCR)方法,用于研究H1N1甲型流感病毒在BHK21细胞中的增殖规律.结果发现:用1MOI H1N1甲型流感病毒接种BHK21,细胞于感染后24 h开始出现细胞病变,36h细胞大量脱落和崩解;病毒定量检测结果显示,H1N1甲型流感病毒增殖迅速,于感染后6、12、24、36 h病毒滴度分别为1.77×106拷贝/mL、2.39×106拷贝/mL、6.58×105拷贝/mL、2.87×105拷贝/mL.研究结果证实本实验建立的RRT-PCR能用于H1N1甲型流感病毒的精确定量,病毒在BHK21细胞中的增殖规律为利用细胞毒生产疫苗提供了参考.

BHK21 cell is a susceptible host for a lot of animal viruses and production. In order to study the replication disciplinarian of influenza A a commonly used cell line for virus vaccines virus in the line of BHK21, the SYBR Green I real-time RT-PCR （RRT-PCR） based on M gene is developed to study the replication disciplinarian of influenza A virus in the line of BHK21. The result shows that cytopathic effect （CPE） is observed at 24 h post infection （P1） when inoculation 1 MOI influenza A virus gets into BHK21 cells, and a large number of cells fall offand the desegregation is at 36 h PI. The virus load is 1.77× 10^6copies/mL at 6 h, 2.39× 10^6copies/mL at 12 h, 6.58× 10^5eopies/mL at 24 h and 2.87× 10^5copies/mL at 36 h PI. The results in this study indicate that the RRT-PCR provides an accurate quantification of influenza A virus. This study also provides basic information for the interaction between cells and virus and vaccine production of influenza A virus.