人呼吸道合胞病毒的稳定性

Stability of Human Respiratory Syncytial Virus

目的观察人呼吸道合胞病毒(HRSV)在不同温度保存、冻融和超声波处理的稳定性。方法将HRSVA2株接种于KMB17细胞上适应培养后,收获病毒液,分别置于37、22、4和-20℃条件下保存不同时间,或经冻融和超声波处理后,用微量细胞病变法检测HRSV的感染性滴度,观察其稳定性。结果HRSV在37℃保存,病毒感染性滴度下降很快,第4天时,降至约1.50lgCCID50/ml,5d后降至1.00lgCCID50/ml以下,几乎检测不到;在22℃保存,病毒感染性滴度下降相对缓慢,第14天时比原始滴度降低约3.20lgCCID50/ml;在4℃和-20℃下保存较稳定,分别保存5周和3个月后,病毒感染性滴度下降<0.60lgCCID50/ml。冻融1、2、3次,病毒感染性滴度分别下降1.37、2.55和3.61lgCCID50/ml,冻融4次后,病毒几乎无感染性。超声波处理后,病毒感染性滴度下降很快,处理1次病毒滴度即下降至原始滴度的50%左右,处理2次后,病毒几乎无感染性。结论HRSV不宜在37℃和22℃存放,可短暂保存于4℃,在-20℃可保存较长时间;冻融和超声波处理均对病毒感染性滴度有明显影响。

Objective To observe the stability of human respiratory syncytial virus（HRSV）to temperature,freeze-thawing and ultrasonication. Methods The KMB17 cells inoculated with HRSV A2 strain were stored at 37,22,4 and -20℃ for various times,or treated by freeze-thawing or ultrasonication,then determined for infectious titer by microcytopathogenic effect assay and observed for stability. Rusults The infectious titer of HRSV was 1.50 lgCCID50 / ml on day 4,while decreased to below 1. 00 lgCCID50 / ml 5 d after storage at 37℃. However,the titer of HRSV decreased by about 3. 20 lgCCID50 / ml on day 14 after storage at 22℃. After storage at 4℃ for 5 weeks or at -20℃ for 3 months,the decrease of HRSV titer was less than 0. 60 lgCCID50 / ml. The HRSV titer decreased by 1. 37,2. 55 and 3.61 lgCCID50 / ml respectively after 1,2 and 3 cycles,and was less than 1. 5 lgCCID50 / ml after 4 cycles of freeze-thawing. The infectious titer of HRSV decreased to about 50% of the original titer after treatment by ultrasonication for 1 time,and was less than 1. 5 lgCCID50 / ml after treatment for 2 times. Conclusion HRSV is unsuitable for storage at 37 and 22℃. However,it may be stored at 4℃ for a short time and at -20℃ for a long time. Both freeze-thawing and ultrasonication influence the infectious titer of HRSV significantly.