ELISA试剂盒检测Sabin株脊髓灰质炎灭活疫苗Vero细胞宿主蛋白残留量的验证  

Verification of a ELISA Kit detection method for residual Vero cell host protein in Sabin strain inactivated poliovirus vaccine

作　　者：程小玲 罗敏 程尧 申瑷琳 郭依依 张娟 李娟娟 施金荣 段凯 CHENG Xiaoling;LUO Min;CHENG Yao;SHEN Ailin;GUO Yiyi;ZHANG Juan;LI Juanjuan;SHI Jinrong;DUAN Kai(Quality Control Department,Wuhan Institute of Biological Products Co.,Ltd.,National Engineering Technology Research Center for Combined Vaccines,Wuhan 430207,Hubei Province,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,湖北武汉430207

出　　处：《微生物学免疫学进展》2023年第5期33-38,共6页Progress In Microbiology and Immunology

基　　金：武汉市科技计划项目(2022023702025187)。

摘　　要：目的验证ELISA试剂盒检测Sabin株脊髓灰质炎灭活疫苗(Sabin strain inactivated poliovirus vaccine,sIPV)中Vero细胞宿主细胞蛋白(host cell protein,HCP)残留量的适用性。方法用同一批ELISA试剂盒检测sIPV中Vero细胞HCP残留量,验证其专属性、重复性、中间精密度、准确度、线性、范围、定量限和耐用性等指标。结果将样品用2种不同稀释液(样品稀释液和疫苗稀释液)稀释后,检测Vero细胞HCP残留量结果均<12.5 ng/mL,表明该方法专属性强;同一检验人员检测同一样品6次,Vero细胞HCP残留量结果均<12.5 ng/mL;不同检验人员检测同一样品6次,Vero细胞HCP残留量结果均<12.5 ng/mL,表明具有良好的重复性和中间精密度;准确度试验中回收率均在99%~106%,CV为2%;在12.5~400.0 ng/mL的线性范围内,标准曲线线性良好(R^(2)>0.98);定量限为50.0 ng/mL;显色时间在25~35 min内对实验无影响,显色温度在36~37℃内对实验无影响,耐用性良好。结论ELISA试剂盒检测sIPV中Vero细胞HCP残留量具有较好的专属性、重复性、中间精密度、准确度、线性和耐用性,可用于sIPV中Vero细胞HCP残留量的检测。Objective To verify the applicability of the ELISA kit for the residual Vero cell host cell protein(HCP)in Sabin strain inactivated poliovirus vaccine(Vero cells).Methods The residual HCP in Sabin strain inactivated poliovirus vaccine(Vero cells)were tested with the same batch of ELISA kits.The specificity,precision(including repeatability and intermediate precision),accuracy,linearity,range,quantitative limit and robustness of the method with kit were verified.Results The detection results of residual protein of Vero cells were all less than 12.5 ng/mL,when diluted with two different solutions,which showed high specificity.The sample was tested by the same tester six times and the result of CV was less than,which showed good repeatability.The same sample was tested by different testers six times and the result of CV was less than,which showed good intermediate precision.The recovery rates in accuracy test were between 99%and 106%and the CV was 2%.The linearity of the standard curves in method were good in the range of 12.5 to 400.0 ng/mL(R^(2)>0.98).The quantitative limit was 50.0 ng/mL.The chromogenic time had no effect on the result of experiment within 25-35 min,and the chromogenic temperature also had no effect on the result of experiment within 36℃-37℃.Therefore the robustness was good.Conclusion The ELISA kit for Vero cell host protein residue in Sabin strain inactivated poliovirus vaccine(Vero cells)showed good specificity,precision(repeatability and intermediate precision),accuracy,linearity and robustness,which was applicable for detection of residual HCP in Sabin strain inactivated poliovirus vaccine(Vero cells).

关 键 词：Sabin株脊髓灰质炎灭活疫苗 VERO细胞 宿主细胞蛋白残留量 酶联免疫吸附试验 验证 

分 类 号：R373.2[医药卫生—病原生物学]