目的 确定使用程序融解系统融解冻存水痘减毒活疫苗收获液的最佳融解温度-时间条件，并分析同一融解温度-时间条件下，不同装量对疫苗原液中的水痘病毒滴度的影响。方法 分别采用程序融解系统在不同融解温度-时间（40 ℃-120 min、50 ℃-90 min、60 ℃-60 min）下融解不同装量（2.5、3.5、4.5 L）的冻存疫苗收获液，然后纯化制备原液，采用噬斑法检测原液病毒滴度，重复3次。选择最佳的融解温度-时间条件，并应用于3批疫苗收获液。结果 不同融解温度-时间和装量条件下融解制备的疫苗原液病毒滴度均符合4.5～6.0 lgPFU/mL的标准。基于Ⅲ类平方和的双因素方差分析显示，温度-时间与装量对疫苗原液病毒滴度的独立主效应差异均具有统计学意义。温度-时间的独立主效应尤为突出(F=31.69，P<0.001)，随着融解温度的升高和时间的缩短，病毒滴度显著增加。装量对病毒滴度的影响差异也具有统计学意义（F=6.75，P=0.006），同一融解温度-时间条件下，随着装量的增加疫苗原液病毒滴度随之下降。融解温度-时间为60 ℃-60 min、装量2.5 L时疫苗收获液制备的原液病毒滴度最高，其均值±标准差为（5.23±0.06） lgPFU/mL。将最佳融解条件应用于3批疫苗收获液，制备的疫苗原液病毒滴度的均值±标准差为（5.27±0.03） lgPFU/mL。结论 疫苗收获液使用程序融解系统的最佳融解温度-时间条件为60 ℃-60 min。同一融解温度-时间条件下，小装量疫苗收获液制备的原液病毒滴度更高。

Objective To determine the optimal thawing temperature-time conditions for thawing frozen live attenuated varicella vaccine harvest by programmed thawing system, and to analyze the influence of different volumes on the virus titer of bulk under the same thawing temperature-time conditions.Methods The frozen vaccine harvest with different volumes (2.5, 3.5, 4.5 L) was thawed under different thawing temperature-time (40 ℃-120 min, 50 ℃-90 min, 60 ℃-60 min) using a programmed thawing system, followed by purification to prepare bulk. The virus titer of bulk was detected by plaque assay, which was repeated 3 times. The optimal thawing temperature-time conditions were selected and applied to 3 batches of vaccine harvest.Results The virus titer of bulk purified from vaccine harvest under different thawing temperature-time and volume all met the standard of 4.5-6.0 lgPFU/mL.Two-way ANOVA based on Class Ⅲ sum of squares showed that the difference between independent main effects of temperature-time and volume on the virus titer of vaccine bulk was statistically significant. The independent main effect of temperature-time was particularly prominent (F = 31.69, P < 0.001), with a significant increase in virus titer at higher thawing temperature and shorter thawing time. The effect of volume on virus titer was also statistically significant (F = 6.75, P = 0.006). Under the same thawing temperature-time condition, the virus titer of vaccine bulk decreased with the increase of volume. When the thawing temperature-time was 60 ℃-60 min and the volume was 2.5 L, the virus titer of the bulk prepared from vaccine harvest was the highest， with mean ± standard deviation of (5.23 ± 0.06) lgPFU/mL. When that optimal thawing condition was applied to 3 batches of vaccine harvest, mean±standard deviation of the virus titer of prepared vaccine bulk was (5.27 ± 0.03) lgPFU/mL.Conclusions The optimal thawing temperature-time condition of vaccine harvest using programmed thawing system is 60 ℃-60 min. Under the same thawing temperature-time condition, the virus titer of bulk prepared from smaller volume of vaccine harvest is higher.