摘要
目的研究人呼吸道合胞病毒(human respiratory syncytial virus,HRSV)在人二倍体KMB17细胞上的增殖动力学,为病毒的体外细胞大量繁殖创造条件.方法将HRSVA2株接种到人二倍体KMB17细胞,采用微量细胞病变法在不同时间检测病毒的感染性滴度,绘制一步生长曲线;同时,以不同剂量的病毒感染复数(MO)I感染细胞,分析细胞开始出现病变及所有接种孔全部病变的时间,并检测收获物的感染性滴度,确定最佳MOI.结果HRSV在KMB17细胞上的感染增殖一步生长曲线分析提示,该病毒在培养到2～4d后开始病变,第5～10天呈对数增长,其增殖高峰为10～11d,10d后增殖趋于平缓;接种MOI越大,开始出现病变时间及完全病变的时间都越早.接种100、10及1个MOI时,接种后2～3d均出现病变,第6天时三组细胞病变率均为100%,感染性滴度为7.37～7.50lgCCID50/mL,三组之间无明显的差异;接种在0.1、0.01及0.001个MOI时,接种后第4天有30%的细胞出现病变,第8～9天时细胞病变率也均达到100%,感染性滴度在6.50～7.0lgCCID50/mL之间;当接种在0.0001个MOI以下时,接种后第5～6天才出现病变,至第14天时细胞仍未能完全产生病变,发生病变的细胞收获物其感染性滴度仅为6.00～6.12lgCCID50/mL.结论HRSVA2株病毒在KMB17细胞内能良好增殖,其增殖高峰为10～11d,收获物感染性滴度最高可达7.50lgCCID50/mL,接种最佳MOI约为0.1～1.0.
Objective To study the growth dynamics of the human respiratory syncytial virus （HRSV） in human diploid cells （KMB17） , to find conditions for the large proliferation of the virus in vitro cells. Methods The A2 strain of HRSV was inoculated into human diploid KMB17 cells. Virus titer at different times was detected by microassay of cytopathic effect, and the growth curves were drawn. At the same time, cells were infected by different multiplicity of infection （MOI）. The time that the first cell and all the inoculated cells lesioned was observed, and the infectious titer of the virus was detected to determine the optimal MOI. Results The results showed that the cell lesion began to appear after virus infected KMB,7 cells 2 - 4 days later. Virus grew in logarithm levels 5 - 10 days later, the peak of virus proliferation was 10 - 11 days. The more dose MOI used, the earlier the cell lesions appeared. Cell began lesion after virus was inoculated 2 - 3 days later, and the cytopathic rate of three groups were 100% 6 days later. The infection titer was 7.37 - 7.50 lg CCIDSO/mL, no significant differences existed among the three groups. When the MOI was 0.1, 0.01 and 0.001, about 30% cell were injured 4 days later. All cells were injured 8 - 9 days later. The infectious titer was 6.50 - 7.00 lg CCID50/mL. When the inoculated dose was less than 0.0001 MOI, the cells injury was found when the virus was inoculated to / cells 5 -6 days later. Part of cells were still normal 14 days later, and the titer was only 6.00 -6.12 lg CCID50/mL. Conclusion The A2 strain of HRSV can grow well in KMB17 cells. The peak of virus proliferation was 10 - 11 days, and the most optimal MOI was 0.1 - 1.0.
出处
《昆明医学院学报》
2009年第8期138-141,共4页
Journal of Kunming Medical College
