A族链球菌核酸检测试剂国家参考品的研制  

Development of national reference materials for group A Streptococcus nucleic acid detection kit

作　　者：石继春[1] 刘茹凤[1] 徐潇[1] 李康[1] 黄洋[1] 李江姣[1] 梁丽[1] 陈驰[1] 龙新星 王春娥[1] 叶强[1] SHI Ji-chun;LIU Ru-feng;XU Xiao;LI Kang;HUANG Yang;LI Jiang-jiao;LIANG Li;CHEN Chi;LONG Xin-xing;WANG Chun-e;YE Qiang(National Institutes for Food and Drug Control,National Center for Medical Culture Collections,Beijing 102629,China)

机构地区：[1]中国食品药品检定研究院中国医学细菌保藏管理中心,北京102629

出　　处：《微生物学免疫学进展》2021年第2期6-12,共7页Progress In Microbiology and Immunology

基　　金：“艾滋病和病毒性肝炎等重大传染病防治科技重大专项”(2018ZX10102-001,2018ZX10734404);国家科技基础条件平台--国家菌种资源库(国家菌种资源库-NMRC-2020-2)。

摘　　要：目的研制A族链球菌核酸检测试剂国家参考品。方法将10株A族链球菌株和10株非A族链球菌参考菌株分别在各自适宜的培养基和温度下培养,收获新鲜培养物进行计数、灭活、稀释和分装,并对参考品进行均匀性和稳定性评估。采用A族链球菌核酸检测试剂对阳性参考品符合率、阴性参考品符合率、重复性和最低检出限进行了适用性验证,并组织4家实验室开展了协作标定。结果建立了由10个阳性参考品、10个阴性参考品、最低检出限参考品和重复性参考品组成的A族链球菌核酸检测试剂评价用国家参考品。经评估该参考品具有良好的均匀性及稳定性,协作标定结果显示阳性参考品符合率、阴性参考品符合率、重复性参考品均符合要求,最低检出限参考品菌液浓度两家实验室标定为1×10^(4)个/mL,两家实验室标定为1×10^(3)个/mL。结论研制的参考品可以用于A族链球菌核酸检测试剂的质量评价。Objective To develop national reference materials for goup A Streptococcus nucleic acid detection kit. Methods 10 strains of group A Streptococcus and 10 strains of non-group A Streptococcus were cultured in a suitable media and temperature, separately. Fresh cultures were harvested, counted, inactivated and subpackaged. The uniformity verification, stability evaluation and accuracy, specificity, repeatability, and minimum detection limit verification of reference materials were conducted, and 4 laboratories were organized to carry out collaborative calibration. Results The national reference materials for group A Streptococcus nucleic acid detection kit were successfully developed, including 10 positive samples, 10 negative samples, minimum detection limit samples and precision samples. They had good uniformity, stability and repeatability in the reference materials. The results of the collaborative calibration showed that the accuracy, specificity and repeatability were in compliance with the regulations, and the minimum detection limit of reference materials was 1.0×10^(3) bacteria/mL in two laboratories, and 1.0×10^(4) bacteria/mL in the other two laboratories. Conclusion The national reference materials for group A Streptococcus nucleic acid detection kit have been verificated and can be used for the quality evaluation of related nucleic acid detection kits in different companies.

关 键 词：A族链球菌 核酸 国家参考品 均匀性 稳定性 协作标定 

分 类 号：R392-33[医药卫生—免疫学]