产气荚膜梭菌核酸检测试剂国家参考品的研制  

Development of a national reference for nucleic acid detection kits of Clostridium perfringens

作　　者：刘茹凤 陈驰 石继春 李康 王斌 王春娥 叶强 李茂光 LIU Rufeng;CHEN Chi;SHI Jichun;LI Kang;WANG Bin;WANG Chune;YE Qiang;LI Maoguang(Division of Bacterial Polysaccharide and Conjugat Vaccines,Institut 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：《国际检验医学杂志》2023年第S01期10-13,共4页International Journal of Laboratory Medicine

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：目的研制产气荚膜梭菌(CP)核酸检测试剂国家参考品。方法通过对5株产气荚膜梭菌和7株非产气荚膜梭菌的菌种检定、培养、灭活及分装,组建产气荚膜梭菌核酸检测试剂国家参考品。对参考品进行样品均匀性以及稳定性评估验证,组织3家实验室协作标定。结果参考品由5份阳性参考品、7份阴性参考品、3份重复性参考品以及1份最低检出限参考品样本组成,均匀性和稳定性良好。协作标定结果显示:5份阳性参考品检测结果均为产气荚膜梭菌阳性;7份阴性参考品检测结果均为产气荚膜梭菌阴性;重复性参考品检测结果全部为气荚膜梭菌阳性(n=10,CV≤5.00%);参考品最低检出限为1.0×10^(3)个菌/mL。结论建立了产气荚膜梭菌核酸检测试剂国家参考品,建立的国家参考品可用于产气荚膜梭菌核酸检测试剂盒的质量控制。Objective To develop a national reference for nucleic acid detection kits of Clostridium perfringens(CP).Methods Five strains of Clostridium perfringens and 7 strains of non-Clostridium perfringens were tested,cultured,inactivated and prepared to establish a national reference for nucleic acid detection kits of Clostridium perfringens.The uniformity and stability of reference were evaluated and verified,and collaborative calibration involving 3 laboratories was organized.Results The reference was composed of 5 positive reference samples,7 negative reference samples,3 repeatable reference samples and 1 minimum detection limit reference sample,with good uniformity and stability.The results of collaborative calibration showed that all the 5 positive reference samples were CP positive,all the 7 negative reference samples were CP negative,the minimum detection limit of reference was 1.0×10^(3) bacterias/mL.The results of repeatable reference were all CP positive(n=10,CV≤5.00%).Conclusion A national reference for nucleic acid detection kits of Clostridium perfringens is established and can be used for quality control of nucleic acid detection kits for Clostridium perfringens.

关 键 词：产气荚膜梭菌 核酸检测 验证 国家参考品 

分 类 号：R446.5[医药卫生—诊断学]