CAJ | 학술논문

目的评估检测百日咳毒素(pertussis toxin,PT)、丝状血凝素(filamentous haemagglutinin,FHA)和黏着素(pertactin,Prn)的3种酶免疫测定试剂盒的稳定性及其在无细胞百日咳疫苗(acellular pertussis vaccine,aPV)生产质量控制中的应用.方法对3种酶免疫测定试剂盒进行热加速试验(37℃放置3 d)和长期稳定性试验(4℃放置6月),根据试剂盒的标准曲线相关系数、最高浓度标准品与阴性对照的吸光度值之比(P/N)值、质控品回收率评估试剂盒的稳定性.将3种试剂盒用于aPV生产的发酵、纯化、吸附阶段的PT、FHA、Prn定量检测,以确定3种试剂盒对aPV生产质量控制的可行性.结果 3种酶免疫测定试剂盒的37℃热加速试验和4℃长期稳定性试验的各项质量参数均符合相关要求.采用试剂盒定量各生产阶段的百日咳抗原含量显示:百日咳杆菌的发酵终点在第42～46 h;各3批PT、FHA和Prn组分液的纯化回收率分别为49.24％、56.12％和63.65％,68.75％、55.60％和49.76％、75.73％、60.63％和50.10％.采用单独吸附的铝佐剂抗原吸附率高于采用混合吸附,但单独和混合吸附方式制备的疫苗的效力无差异.结论检测PT、FHA和Prn的3种酶免疫测定试剂盒具有良好的稳定性,可用于aPV生产的质量控制.
목적 평고검측백일해독소(pertussis toxin,PT)、사상혈응소(filamentous haemagglutinin,FHA)화점착소(pertactin,Prn)적3충매면역측정시제합적은정성급기재무세포백일해역묘(acellular pertussis vaccine,aPV)생산질량공제중적응용.방법 대3충매면역측정시제합진행열가속시험(37℃방치3 d)화장기은정성시험(4℃방치6월),근거시제합적표준곡선상관계수、최고농도표준품여음성대조적흡광도치지비(P/N)치、질공품회수솔평고시제합적은정성.장3충시제합용우aPV생산적발효、순화、흡부계단적PT、FHA、Prn정량검측,이학정3충시제합대aPV생산질량공제적가행성.결과 3충매면역측정시제합적37℃열가속시험화4℃장기은정성시험적각항질량삼수균부합상관요구.채용시제합정량각생산계단적백일해항원함량현시:백일해간균적발효종점재제42～46 h;각3비PT、FHA화Prn조분액적순화회수솔분별위49.24％、56.12％화63.65％,68.75％、55.60％화49.76％、75.73％、60.63％화50.10％.채용단독흡부적려좌제항원흡부솔고우채용혼합흡부,단단독화혼합흡부방식제비적역묘적효력무차이.결론 검측PT、FHA화Prn적3충매면역측정시제합구유량호적은정성,가용우aPV생산적질량공제.
Objective To evaluate the stability of 3 kinds of EIA kits for detecting pertussis toxin (PT),filamentous haemagglutinin (FHA) and pertactin (Prn) and their application in the quality control of acellular pertussis vaccine (aPV) production.Methods The stability of EIA kits were evaluated by heat accelerated test (37 ℃ for 3 days) and long-term stability test (4 ℃ for 6 months),using the correlation coefficient of standard curve,the P/N value and the recovery of the quality control material.The contents of PT,FHA and Prn in the fermentation,purification and adsorption stages during the production process of aPV were detected by these 3 kinds of EIA kits,and the feasibility of using them in the quality control of aPV production were determined.Results Heat accelerated stability test and long-term stability test showed that quality parameters of 3 kinds of EIA kits all met the requirement.The detection results of pertussis antigens in the different production stages of pertussis vaccine obtained by 3 kinds of EIA kits showed that the fermentation end of Bordetella pertussis was at 42-46 h,and the purification recovery rates were 49.24％,56.12％ and 63.65％ for 3 lots of PT antigen,respectively,68.75％,55.60％ and 49.76％ for 3 lots of FHA antigen,respectively,and 75.73％,60.63％ and 50.10％ for 3 lots of Prn antigen,respectively.The antigen absorption rates of aluminum adjuvant with separate absorption were higher than those with mixed adsorption,but there was no difference in potency between both vaccines prepared by separate and mixed adsorption.Conclusion Three kinds of EIA kits for detecting PT,FHA and Prn have good stability and can be used for the quality control of aPV production.