细胞密度和感染复数对SARS病毒在Vero细胞中增殖的影响

Optimal Cell Density and Multiplicity of Infection for the Propagation of SARS-CoV in Vero Cells

阐明宿主细胞密度和病毒感染复数(MOI)对SARS冠状病毒增殖的影响,为SARS灭活疫苗的研究奠定基础。采用析因设计方法,考虑MOI(0.01、0.05和0.1)与细胞密度(2×104、4×104和8×104/cm2)2个3水平实验因素,在各水平组合条件下培养SARS病毒,以组织培养半数感染量(TCID50)作为观测指标,分析SARS冠状病毒滴度在不同增殖条件下的差异。结果表明,MOI、宿主细胞密度和二者之间的交互作用对SARS病毒滴度的影响分别具有非常显著性差异(F=12.70,P<0.01)、显著性差异(F=6.94,P<0.05)和非常显著性差异(F=8.22,P<0.01)。在0.01MOI和8×104/cm2条件下,病毒滴度达5.57×105TCID50/mL。说明SARS冠状病毒在培养细胞中的增殖能力显著依赖于宿主细胞密度、病毒感染复数和二者的交互作用。

To clarify the effects of cell density at infection and multiplicity of infection (MOI) on SARS coronavirus propagation in Vero cell cultures and lay a foundation for SARS-CoV vaccine development. The factorial design methods and analysis of variance were adopted to compare SARS-CoV titers (TCID50) after infection at MOIs of 0.01, 0.05 and 0.1 with different cell density of 2×104, 4×104 and 8×104 cell/cm2. There was great significant difference in the mean TCID50 among MOI of 0.01, 0.05 and 0.1(F=12.70, P<0.01). There was significant difference in the mean TCID50 among 2×104, 4×104 and 8×104 cells/cm2 (F=6.94, P<0.05). There was great significant difference in the mean TCID50 in reciprocal condition(F=8.22, P<0.01). The optimal SARS-CoV titer (5.57×105 TCID50/mL) was obtained with cultures seeded at about 8×104 cells /cm2 and at an MOI of 0.01. These results suggested that SARS coronavirus propagation dependent upon cell density, MOI and their reciprocity at infection.