口服六价重配轮状病毒减毒活疫苗(Vero细胞)病毒滴度工作参考品的研制  

Establishment and stability studies of a viral titer reference for oral hexavalent reconstituted live attenuated rotavirus vaccine(Vero cell)

作　　者：邹文琪 王梅珂 周蓉 程小玲 姜志军 白萱 石晨 郭旭梦 李榆杨 孙露 钟守红 石艳红 何亮 申瑷琳 罗敏 Zou Wenqi;Wang Meike;Zhou Rong;Cheng Xiaoling;Jiang Zhijun;Bai Xuan;Shi Chen;Guo Xumeng;Li Yuyang;Sun Lu;Zhong Shouhong;Shi Yanhong;He Liang;Shen Ailin;Luo Min(Department of Quality Control,Wuhan Institute of Biological Products Co.,Ltd.,National Engineering Technology Research Center for Combined Vaccines,Wuhan 430207,China;Department of Rotavirus Vaccine,Wuhan Institute of Biological Products Co.,Ltd.,National Engineering Technology Research Center for Combined Vaccines,Wuhan 430207,China;Department of Quality Assurance,Wuhan Institute of Biological Products Co.,Ltd.,National Engineering Technology Research Center for Combined Vaccines,Wuhan 430207,China)

机构地区：[1]武汉生物制品研究所有限责任公司质量控制室,国家联合疫苗工程技术研究中心,武汉430207 [2]武汉生物制品研究所有限责任公司轮状病毒疫苗室,国家联合疫苗工程技术研究中心,武汉430207 [3]武汉生物制品研究所有限责任公司质量保证部,国家联合疫苗工程技术研究中心,武汉430207

出　　处：《国际生物制品学杂志》2025年第1期32-38,共7页International Journal of Biologicals

摘　　要：目的建立口服六价重配轮状病毒减毒活疫苗(Vero细胞)病毒滴度工作参考品,标定工作参考品范围并考察其稳定性。方法采用Ⅲ期临床试验用批次原液制备6个血清型病毒滴度工作参考品,使用荧光灶试验标定各血清型工作参考品范围,研究新旧参考品的同质性,并考察各血清型参考品在不同条件的稳定性。结果建立的轮状病毒G1、G2、G3、G4、G8、G9血清型工作参考品的病毒滴度标定范围分别为7.5~8.7、6.4~7.8、6.6~7.8、6.4~7.6、7.1~8.3、7.2~8.4 lg荧光灶单位/mL。用新旧参考品进行滴度测定的同一样品,各血清型滴度结果变异系数<5%且差异无统计学意义。各血清型轮状病毒工作参考品在2~8℃和25℃条件下分别储存至7 d稳定性良好;在37℃条件下,工作参考品可以储存3~7 d;在冻融2次条件下,参考品稳定性良好;在长期稳定性考察中,-60℃储存至18个月稳定性良好,实验间变异系数<5%,病毒滴度均在标定范围内。结论建立并确认标定范围的口服六价重配轮状病毒减毒活疫苗(Vero细胞)病毒滴度工作参考品,在不同条件下具有良好的储存稳定性,能够保证检测结果的一致性。Objective To establish reference substance for virus titer detection of oral hexavalent reconstituted rotavirus attenuated live vaccines(Vero cell),and to determine the range and stability of the reference.Methods Reference substances for virus titer detection of 6 rotavirus serotypes were prepared with rotavirus stock solution validated in phaseⅢclinical trials.A comprehensive evaluation of new and old reference substances was conducted using fluorescence focus assay,and the stability of each serotype reference under different conditions was monitored.Results The viral titer calibration ranges for serotypes G1,G2,G3,G4,G8,and G9 rotavirus working references were 7.5-8.7,6.4-7.8,6.6-7.8,6.4-7.6,7.1-8.3,and 7.2-8.4 lg fluorescent focus units/mL,respectively.The coefficients of variation(CVs)of the titer results for each serotype of the same sample were<5%and not statistically significantly different when titered against the old and new references.The working reference samples for all 6 serotypes of rotavirus had good stability when stored at 2-8℃and 25℃for 1,3,7 days,or after 2 freeze-thaw cycles.Storage at 37℃for 3-7 days had no impact on the virus titer.The stability was good after storage at-60℃for 12 months with CV between tests<5%.Conclusion The established and range-determined reference material for rotavirus vaccine titer has good stability under different conditions and can ensure the consistency of test results.

关 键 词：轮状病毒 参考品 稳定性 疫苗 减毒 

分 类 号：R392-33[医药卫生—免疫学]