肺炎克雷伯菌核酸检测试剂国家参考品的研制  

Development of national reference product for nucleic acid detection of Klebsiella pneumoniae

作　　者：刘茹凤 李江姣 石继春 李康 黄洋 王斌 王春娥 叶强 LIU Rufeng;LI Jiangjiao;SHI Jichun;LI Kang;HUANG Yang;WANG Bin;WANG Chune;YE Qiang(Division of Bacterial Polysaccharide and Conjugate Vaccines,Institute for Biological Product Control,National Institutes for Food and Drug Control,Beijing,China,102629)

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：《分子诊断与治疗杂志》2025年第1期5-8,12,共5页Journal of Molecular Diagnostics and Therapy

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：目的研制肺炎克雷伯菌(KP)核酸检测国家参考品。方法通过对10株肺炎克雷伯菌和9株非肺炎克雷伯菌的菌种检定、培养、灭活及分装,建立肺炎克雷伯菌核酸检测试剂国家参考品。对参考品进行均匀性以及稳定性评估验证,组织8家实验室进行协作标定。结果参考品由10份阳性、9份阴性、3份精密性以及1份最低检出限样品组成,均匀性和稳定性良好。协作标定结果显示:10份阳性参考品检测结果均为阳性;9份阴性参考品:除一家企业的N7不符合要求其余均为阴性;最低检出量参考品的检出量不高于1.0×10^(3)个菌/mL;重复性参考品检测结果一致(n=10,CV≤5%)。结论建立的肺炎克雷伯菌核酸检测试剂国家参考品填补了相关领域的空白,能够用于肺炎克雷伯菌核酸检测试剂的质量控制。Objective To develop and validate the national reference for nucleic acid detection of Klebsiella pneumoniae(KP).Methods Through the identification,culture,inactivation and packaging of 10 strains of Klebsiella pneumoniae and 9 strains of non⁃Klebsiella pneumoniae,the national reference for nucleic acid testing of Klebsiella pneumoniae was established.The uniformity and stability of reference samples were evaluated and verified,and 9 laboratories were organized to coordinate calibration.Results The reference samples consisted of 10 positive samples,9 negative samples,3 precision samples and 1 minimum detection limit sample,all exhibiting good uniformity and stability.The collaborative calibration results indicated that all 10 positive reference samples tested positive.Of the 9 negative reference samples,only N7 tested positive when using the test kit from one enterprise,failing to meet the requirements.The minimum detection limit for the ref⁃erence substance should not exceed 1.0×10^(3) bacteria/mL.Repeated testing of reference samples should yield con⁃sistent results(n=10,CV≤5%).Conclusion The established national reference for detecting Klebsiella pneu⁃moniae nucleic acid fills a gap in the field and can be used for quality control of related reagents.

关 键 词：肺炎克雷伯菌 核酸检测 验证 国家参考品 

分 类 号：R943[医药卫生—药剂学]