人偏肺病毒标准株在不同细胞中的复制动力学

Replication Kinetics of Standard Human Metapneumovirus Strain in Various Cell Lines

目的探讨人偏肺病毒(Human metapneumovirus,hMPV)标准株在不同细胞中的复制动力学,确定适合分离和培养hMPV的细胞。方法将hMPV A亚型标准株hMPV/NL/1/00和B亚型标准株hMPV/NL/1/99分别接种于Vero、Vero-E6、LLC-MK2、A549和HEp-2细胞,盲传数代,逐日观察细胞病变(CPE),并于接种后第2、4、6、8、10、12、14、16、18、20天提取细胞RNA,逆转录合成cDNA,采用实时荧光定量PCR检测hMPV F蛋白基因。结果接种hMPV后3 d可在Vero、Vero-E6、LLC-MK2和A549细胞中观察到CPE,不同亚型的hMPV所致的CPE无差别;hMPV可在Vero、Vero-E6和LLC-MK2细胞中稳定复制,不能在A549和HEp-2细胞中稳定传代。结论 Vero、Vero-E6和LLC-MK2细胞是适合培养hMPV的细胞,A549和HEp-2细胞不适合用于培养hMPV。

Objective To investigate the replication kinetics of standard human metapneumovirus（hMPV） strain in various cell lines to find a cell line suitable for the isolation and culture of the virus.Methods Standard hMPV strain hMPV / NL / 1 / 00 of subtype A and hMPV / NL / 1 / 99 of subtype B were inoculated to Vero,Vero-E6,LLC-MK2,A549 and HEp-2 cells separately for several blind passages.The CPEs were observed daily,and the RNAs were extracted from the cells on days 2,4,6,8,10,12,14,16,18 and 20 after inoculation,then reversely transcribed to cDNA and determined by real-time fluorescent quantitative PCR.Results CPEs were observed in Vero,Vero-E6,LLC-MK2 and A549 cells 3 d after inoculation.However,the CPEs caused by hMPV strains of various subtypes showed no significant difference.The hMPV could be stably replicated in Vero,Vero-E6 and LLC-MK2 cells,but could not be subcultured stably in A549 and HEp-2 cells.Conclusion Vero,Vero-E6 and LLC-MK2 cells were suitable,while A549 and HEp-2 cells were unsuitable for the culture of hMPV.