HPA基因分型国家参考盘的定值和验证  

Collaborative calibration and verification of a national reference panel for HPA genotyping

作　　者：胡泽斌[1] 王红梅[2] 孙彬裕[1] 段生宝[2] 张河战[1] HU Zebin;WANG Hongmei;SUN Binyu;DUAN Shengbao;ZHANG Hezhan(National Institutes for Food and Drug Control,NMPA Key Laboratory for Quality Research and Evalua-tion of In vitro Diagnostic Reagents,Beijing,China,100050;Suzhou Institute of Biomedical Engineering and Technology(SIBET),Chinese Academy of Sciences(CAS),Suzhou,Jiangsu,China,215163)

机构地区：[1]中国食品药品检定研究院,国家药品监督管理局体外诊断试剂质量研究与评价重点实验室,北京100050 [2]中国科学院苏州生物医学工程技术研究所,江苏苏州215163

出　　处：《分子诊断与治疗杂志》2024年第11期2034-2037,共4页Journal of Molecular Diagnostics and Therapy

基　　金：国家重点研发计划(2023YFC2413100)。

摘　　要：目的对HPA基因分型首批国家参考盘进行准确定值和进一步均匀性、稳定性验证。方法选择具有良好资质的5家实验室按统一的协作标定方案进行联合定值,采用PCR⁃SSP法、PCR⁃荧光探针法、荧光PCR染料法、荧光PCR熔解曲线法、PCR⁃SBT测序法、NGS测序法对HPA基因分型首批国家参考盘进行协作标定;采用PCR⁃SSP法、荧光PCR法和SBT测序法,考察HPA基因分型首批国家参考盘在量值水平的稳定性和均匀性。结果5家实验室多种方法学联合协作标定的结果完全一致,对HPA国家参考盘样本进行了HPA⁃1~29抗原基因分型结果的准确定值。该参考盘瓶间和瓶内均匀性良好、冻融稳定性良好。结论HPA基因分型首批国家参考盘的量值准确性、均匀性和稳定性均符合要求,可用于基于PCR方法的多种不同检测范围的HPA分型检测试剂盒的产品质量评价。Objective To accurately determine the values and further verify the uniformity and stability of the first batch of national reference panel for HPA genotyping.Methods Five laboratories with strong qualifications were chosen to participate in a joint calibration process based on a unified collaborative calibration plan.The PCR⁃SSP method,PCR fluorescence probe method,fluorescence PCR dye method,fluorescence PCR melting curve method,PCR⁃SBT sequencing method,and NGS sequencing method were all used for this collaborative calibration effort.The stability and uniformity of the HPA panel were examined at a quantitative level using the PCR SSP method,fluorescence PCR method,and SBT sequencing method.Result The results of the collaborative calibration of multiple methodologies by 5 laboratories were completely consistent.The HPA⁃1⁃29 antigen genotyping results were accurately determined for the HPA national reference panel.The reference panel exhibited good uniformity both between and within the bottles,as well as good freeze⁃thaw stability.The results were entirely consistent with the typing results of normal samples stored at-20℃.Conclusion The accuracy,uniformity,and stability of the HPA genotyping national reference panel meet the necessary requirements.It can be used for evaluating the product quality of various HPA genotyping detection kits that are based on PCR methods,each with different detection ranges.

关 键 词：HPA 协作标定 准确性 质量评价 体外诊断试剂 

分 类 号：R927.1[医药卫生—药学]