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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) stored at different temperatures, freeze-thaw and sonication. Methods After HRSVA2 strain was inoculated on KMB17 cells and cultured, the virus solution was harvested and stored at 37, 22, 4 and -20 ℃ for different time or frozen-thawed and sonicated, respectively. Infectious titers of HRSV were observed for their stability. Results HRSV was stored at 37 ° C and the titer of viral infectivity dropped rapidly. On the 4th day, the titer decreased to about 1.50lgCCID50 / ml and dropped below 1.00lgCCID50 / ml after 5 days. At 22 ℃, the virus was infected The titer decreased relatively slowly, about 3.20lgCCID50 / ml lower than the original titer on the 14th day, and was more stable at 4 ℃ and -20 ℃. After 5 weeks and 3 months respectively, the virus titer decreased < 0.60 lg CCID50 / ml. 1, 2 and 3 freeze-thaw cycles, the viral infectivity titers decreased by 1.37, 2.55 and 3.61lgCCID50 / ml, respectively. After freeze-thaw four times, the virus was almost non-infectious. After the ultrasonic treatment, the viral infectious titer dropped rapidly, and the virus titer decreased to about 50% of the original titer after the treatment of one virus. After being treated twice, the virus was almost non-infectious. Conclusion HRSV is not suitable for storage at 37 ℃ and 22 ℃, but can be stored at 4 ℃ for a short time and stored at -20 ℃ for a long time. Both freeze-thaw and ultrasonic treatment have a significant effect on viral titer.