肺炎链球菌核酸检测试剂国家参考品的研制及验证  

Preparation and validation of a national reference for nucleic acid test reagent of Streptococcus pneumoniae

作　　者：李康 黄洋 王春娥 李江姣 梁丽 陈驰 刘茹凤 叶强 LI Kang;HUANG Yang;WANG Chun’e;LI Jiangjiao;LIANG Li;CHEN Chi;LIU Rufeng;YE Qiang(Division of Bacterial Polysaccharide and Conjugate Vaccines,Institute for Biological Product Control,National Institutes for Food and Drug Control,Beijing 102629,China)

机构地区：[1]中国食品药品检定研究院生物制品检定所细菌多糖和结合疫苗室,北京102629

出　　处：《中国医药导报》2021年第15期141-144,共4页China Medical Herald

基　　金：国家科技重大专项课题(2018ZX10102-001-002、2018ZX09738006-006);国家科技基础条件平台—国家菌种资源库课题(NMRC-2020-2)。

摘　　要：目的研制肺炎链球菌核酸检测试剂国家参考品,并进行适用性验证。方法选择10株肺炎链球菌和10株非肺炎链球菌细菌制备成阳性参考品、阴性参考品、重复性参考品和最低检出限参考品。对参考品的分装均匀性和稳定性进行评估。采用4家企业生产的肺炎链球菌核酸检测试剂盒对制备的肺炎链球菌国家参考品进行准确性、特异性、重复性和最低检出限验证。结果参考品的分装均匀度良好,循环阈值的变异系数均在5.00%以内。2~8℃、室温(25℃)和37℃放置3、7 d及反复冻融3、5次都不影响参考品的稳定性。4家企业的试剂盒的准确性、特异性及重复性检测结果均符合要求。1家企业的试剂盒的最低检出限为1.0×104个/mL,其他3家企业的试剂盒的最低检出限为1.0×103个/mL。结论研制完成一套肺炎链球菌核酸检测试剂国家参考品,能够用于肺炎链球菌核酸检测试剂盒的质量控制和评价。Objective To prepare a national reference for nucleic acid test reagent of Streptococcus pneumoniae and validate its applicability.Methods Ten strains of Streptococcus pneumoniae and ten strains of non-Streptococcus pneumoniae were selected to prepare positive reference,negative reference,repeatable reference and minimum detection limit reference.The uniformity and stability of the reference package was evaluated.The accuracy,specificity,reproducibility and minimum detection limit of the prepared national reference of Streptococcus pneumoniae were validated by using nucleic acid detection kits of Streptococcus pneumoniae produced by four enterprises.Results The packing uniformity of the reference was good,and the coefficient of variation of the cycle threshold values were all within 5.00%.The stability of the reference was not affected by being placed for three or seven days at 2-8℃,room temperature(25℃)and 37℃and repeated freezing-thawing for three or five times.The accuracy,specificity and reproducibility of the kits from the four enterprises were all in line with the requirements.The minimum detection limit of the kits from one enterprise was 1.0×104/mL,and the minimum detection limit of the kits from the other three enterprises was 1.0×103/mL.Conclusion A set of national reference reagents for nucleic acid test reagent of Streptococcus pneumoniae has been prepared,which can be used for the quality control and evaluation of Streptococcus pneumoniae nucleic acid detection kits.

关 键 词：肺炎链球菌 核酸检测试剂 国家参考品 质量评价 

分 类 号：R378.1[医药卫生—病原生物学]