狂犬病疫苗效价测定改良NIH法建立及应用  

Establishment and application of M-NIH for potency test of rabies vaccine

作　　者：吴小红[1] 曹守春[1] 石磊泰[1] 王云鹏[1] 赵丹华 李加[1] 李玉华[1] WU Xiaohong;CAO Shouchun;SHI Leitai;WANG Yunpeng;ZHAO Danhua;LI Jia;LI Yuhua(NMPA Key Laboratory for Quality Control and Evaluation of Biological Products,Division of Arbovirus Vaccine,National Institute for Food and Drug Control,Beijing 102629,China)

机构地区：[1]中国食品药品检定研究院虫媒病毒疫苗室,国家药品监督管理局生物制品质量研究与评价重点实验室,北京102629

出　　处：《药学研究》2023年第10期790-797,共8页Journal of Pharmaceutical Research

基　　金：国家科技重大专项重大新药创制(No.2018ZX09738005)。

摘　　要：目的按照动物试验3R原则,建立改良NIH法(modified NIH,M-NIH)即单一稀释法检测狂犬病疫苗效价,替代传统的NIH法。方法通过比较单一稀释度的待检疫苗与第八批狂犬病疫苗效力试验用国家标准品的小鼠保护率,建立狂犬病疫苗效价测定的改良NIH法,并对该方法进行重复性、准确性及适用性验证。采用M-NIH法和NIH法对52批狂犬病疫苗进行比较研究,再联合9家实验室验证共计247批次,使用Kappa系数和Mcnemar-Bowker检验评价两种方法的一致性。并采用3家企业生产的狂犬病疫苗,进行适用性验证。对M-NIH法获批准后近4年来应用于狂犬病疫苗批签发检验做回顾性分析。结果52批次狂犬病疫苗效价测定的两种方法比较研究结果显示,两种方法的符合率为92.3%(Kappa=0.82,P<0.01);247批次扩大验证结果表明两种方法的符合率为80.2%(Kappa=0.54,P<0.01)。采用M-NIH法测定标准品保护率时,实验室内变异系数为21.7%,实验室间变异系数为23.4%。3家企业批签发狂犬病疫苗适用性验证结果表明M-NIH重复性检测结果与NIH结果符合率为100%。近4年来批签发机构采用M-NIH法抽检820批狂犬病疫苗效价,合格率为93.8%;M-NIH不合格批次最终经NIH法测定后,合格率合计为99.8%。结论建立了改良NIH法。该方法获得批准后用于狂犬病疫苗的效价检测,大大减少了动物使用量,提高了狂犬病疫苗的批签发效率。Objective To establish a modified NIH(M-NIH,single dilution method)to test the potency of rabies vaccine according to the 3R principles of animal testing,and to replace the traditional NIH method.Methods Modified NIH(M-NIH)method was established by comparing the protection rate of mice between the single dilution of rabies vaccine and the eighth batch of national standard reference used in the potency test of rabies vaccine.And the repeatability,accuracy and applicability of M-NIH were verified.52 batches of rabies vaccines were compared by M-NIH method and NIH method,and a total of 247 batches of rabies vaccines were verified by 9 laboratories.Kappa coefficient and Bowker test were used to evaluate the consistency of the two methods.The applicability of rabies vaccines from three manufactures was verified.A retrospective analysis in the past 4 years was made on the application of rabies vaccine lot release using M-NIH method after approval.Results The verification results of 52 batches of rabies vaccine showed that the coincidence rate of the two methods was 92.3%(Kappa=0.82,P<0.01),and the coincidence rate of the 247 batches rabies vaccines was 80.2%(Kappa=0.54,P<0.01)in the extended verification.The coefficient of variation of M-NIH standard protection rate was 21.7%in intra-laboratory and 23.4%in inter-laboratory.The applicability verification results of three manufactures showed that the coincidence rate of M-NIH results and NIH results was 100%.In the past four years,a total of 820 batches of rabies vaccines were tested,and the qualified rate was 93.8%by using M-NIH,M-NIH unqualified batches were finally determined by NIH method,and the qualified rate of rabies vaccine was 99.8%.Conclusion M-NIH was established and approved to be used to test the potency of rabies vaccine.In the past 4 years,the efficiency of lot release of rabies vaccine was highly improved and the use of experiment mouse was greatly reduced by using M-NIH.

关 键 词：人用狂犬病疫苗 效价测定NIH法 改良NIH法 方法学验证 

分 类 号：R917[医药卫生—药物分析学]