组分百日咳疫苗安全性检测体外替代方法的应用和初步评价  

Application and Preliminary Evaluation of in Vitro Alternative Methods for the Safety Testing of Component Pertussis Vaccine

作　　者：吴燕[1] 卫辰[1] 王丽婵 晁哲[1] 马霄[1] Wu Yan;Wei Chen;Wang Lichan;Chao Zhe;Ma Xiao(National Institutes for Food and Drug Control,Key laboratory of the Ministry of Health for Research on Quality and Standardization of Biotech Products,Beijing 102629,China)

机构地区：[1]中国食品药品检定研究院卫生部生物技术产品检定方法及其标准化重点实验室,北京102629

出　　处：《中国药事》2023年第9期1047-1053,共7页Chinese Pharmaceutical Affairs

摘　　要：目的:用一种组分百日咳疫苗残留毒性的体外替代检测方法,更稳定和客观地评价疫苗成品的生产一致性。方法:验证直接定性法和间接定量法2种中华仓鼠卵巢细胞簇集试验方法,分别对6个国内外厂家的10种组分百日咳疫苗产品毒性进行定量和定性检测,并使用百日咳毒素参考品将体外法与小鼠组胺致敏试验进行桥接和初步评价。结果:体外定量试验的灵敏度为0.0026±0.0003 IU·mL^(-1),几何变异系数为13%,体外定性试验的灵敏度为0.0067±0.0016 IU·mL^(-1),几何变异系数为24%,组胺致敏试验的灵敏度为3.3 IU·mL^(-1);所有样品的体外定性结果均为阴性,小鼠组胺致敏试验结果均合格,体外定量结果与小鼠组胺致敏结果具有良好的相关性(r=0.737,P<0.05)。结论:本研究在国内首次使用CHO细胞簇集试验方法检测百日咳疫苗成品,并进行了初步评价。该方法灵敏度高于小鼠组胺致敏试验,可应用于百日咳疫苗毒性及毒性逆转检测。该方法需要更广泛的推广应用和数据积累,以达到完全替代动物实验的目标。Objective:To use an in vitro alternative detection method for residual toxicity of component pertussis vaccine,and to evaluate the consistency of production of vaccine products more stably and objectively.Methods:Two CHO cell clustering assays were established and validated by direct qualitative method and indirect quantitative method.The toxicity of ten component of pertussis vaccine products from six domestic and foreign manufacturers were tested quantitatively and qualitatively,the in vitro method and histamine sensitization test of mice were bridged and evaluated using pertussis toxin reference.Results:The sensitivity of in vitro quantitative test was 0.0026±0.0003 IU·mL^(-1),GCV was 13%,the sensitivity of in vitro qualitative test was 0.0067±0.0016 IU·mL^(-1),GCV was 24%,and the sensitivity of the histamine sensitization test was 3.3 IU·mL^(-1);the qualitative results of all samples in vitro were negative,the results of histamine sensitization test in mice were all qualified,and there was a good correlation between the quantitative results in vitro and histamine sensitization results in mice(r=0.737,P<0.05).Conclusion:In this study,CHO cell cluster assay was used to detect pertussis vaccine for the first time in China and conducted a preliminary evaluation.The sensitivity of this method is higher than that of histamine sensitization test in mice,and it could be used to detect the toxicity and toxicity reversal of pertussis vaccine.This method requires a wider range of applications and accumulation of data to achieve the goal of completely replacing animal experiments.

关 键 词：组分疫苗 百日咳 中华仓鼠卵巢细胞簇集试验 毒性残留 替代方法 

分 类 号：R95[医药卫生—药学]