摘要目的:建立Sabin株脊髓灰质炎灭活疫苗（Vero细胞）病毒灭活验证方法，并进行方法学验证。方法:将Ⅰ、Ⅱ、Ⅲ型脊髓灰质炎病毒进行系列梯度稀释，分别接种于Vero细胞，进行2次盲传扩增，观察细胞病变情况。对该方法的最低检测限及细胞对病毒的敏感度进行摸索，并对该方法的精密度（重复性和中间精密度）和适用性进行验证。结果:建立的病毒灭活验证方法对Ⅰ、Ⅱ、Ⅲ型脊髓灰质炎病毒的检测限分别为-2、-1、0 lgCCID 50/mL。Ⅰ、Ⅱ、Ⅲ型脊髓灰质炎病毒对Vero细胞的最小接种浓度均为2 lgCCID 50/mL，在第7天均可观察到Vero细胞病变，即该细胞的敏感度为2 lgCCID 50/mL。同一检验人员重复检测同一批样品6次结果一致；2名检验人员在不同时间重复检测同一批样品6次结果一致；样品在-70 ℃及以下存放7 d与未冻存的样品检验结果一致；分别对生产过程中产生的Sabin株脊髓灰质炎灭活疫苗（Vero细胞）Ⅰ、Ⅱ、Ⅲ型单价灭活病毒液及原液各3批进行病毒灭活验证，取样当天和在-70 ℃及以下存放7 d后检验的样品的灭活验证检验结果一致，表明该方法的精密度和适用性均较好。 结论:Sabin株脊髓灰质炎灭活疫苗（Vero细胞）病毒灭活验证方法能有效检测样品的灭活状态，可用于对病毒灭活效果的评价及工艺过程中对中间品等关键步骤的监控。

abstractsObjective:To establish and verify the verification method of virus inactivation of Sabin strain inactivated poliovirus vaccine (Vero cell)(sIPV).Methods:Poliovirus types Ⅰ, Ⅱ and Ⅲ were inoculated into Vero cells by a series of gradient dilution, and then amplified by blind cell passage twice. The cytopathic changes were observed, the minimum detection limit of this method and cell sensitivity to virus were explored, and the precision (repeatability and intermediate precision) and applicability of this method were verified.Result:The detection limits of the established virus inactivation validation method for poliovirus types Ⅰ, Ⅱ and Ⅲ were -2, -1 and 0 lgCCID 50/mL, respectively. The minimum inoculation concentration of poliovirus types Ⅰ, Ⅱ and Ⅲ to Vero cells was 2 lgCCID 50/mL, and the Vero cell lesions were all observed on day 7 .Thus, the sensitivity of the cell was 2 lgCCID 50/mL. The same inspector repeatedly tested the same batch of samples 6 times, and the test results were consistent. The same batch of samples were tested 6 times by different inspectors at different time, and the test results were consistent. The result of sample stored at -70 °C or below for 7 d was consistent with that of unfrozen sample.The inactivation verification test was carried out on 3 batches of types Ⅰ, Ⅱ and Ⅲ monovalent inactivated virus solution and bulk from sIPV production process, and the inactivation verification test results of the samples tested on the day of sampling and after 7 d of storage at -70 °C or below were consistent, indicating that the precision and applicability of the method were good. Conclusion:The sIPV virus inactivation validation method effectively detects the inactivation status of the sample, which can be used to evaluate the virus inactivation effect and monitor key steps such as intermediates in the process.