粪肠球菌核酸检测试剂国家参考品的研制

Development of national reference materials for Enterococcus faecallis nucleic acid detection kit

中文关键词:  粪肠球菌  核酸  国家参考品  均匀性  稳定性  协作标定

英文关键词:Enterococcus faecalis  nucleic acid  national reference materials  uniformity  stability  collaborative study

中文摘要:

      目的 研制粪肠球菌核酸检测试剂的国家参考品。方法 将9株粪肠球菌和8株非粪肠球菌参考品候选菌株分别在各自适宜的培养条件下培养, 收集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。组织3家实验室对研制的参考品进行了协作标定。结果 研制了由9份阳性候选参考品、8份阴性候选参考品、重复性参考品和最低检出限参考品组成的粪肠球菌参考品。阳性参考品、最低检出限参考品和重复性参考品循环阈值(cycle threshold, Ct)的变异系数(coefficient of variation, CV)均在5.0%以内, 分装均匀性良好; 参考品各成分的稳定性良好(P值均小于0.05); 协作标定结果显示阳性参考品符合率及阴性参考品符合率均为100%, 重复性参考品检测Ct值的CV均在5.0%以内, 参考品最低检出限为1.0×103个/mL。结论 研制的参考品可用于粪肠球菌核酸检测试剂的质量评价。

英文摘要:

      Objective To develop national reference materials for Enterococcus faecalis nucleic acid detection kit. Methods Nine strains of Enterococcus faecalis and 8 candidate strains of non-Enterococcus faecalis reference samples were cultured under their respective suitable culture conditions. Fresh cultures were collected for counting, inactivation, dilution and packing, and the reference samples were evaluated for homogeneity and stability. Three laboratories were organized to calibrate the developed reference. Results The coefficient of variation (CV) of cycle threshold (Ct) of positive reference, minimum detection limit reference and repeatable reference were all less than 5.0%, and the distribution uniformity was good. The stability of the reference components was good (P<0.05); the cooperative calibration results showed that the coincidence rate of positive reference and negative reference were 100%, the Ct value of repeatable reference was less than 5.0%, and the minimum limit of detection reference was 1×103/mL. An Enterococcus faecalis reference product was prepared, which consisted of 9 positive candidate reference products, 8 negative candidate reference products, repeatable reference products and minimum detection limit reference products. Conclusion The developed reference material can be used for the quality evaluation of nucleic acid detection reagent for Enterococcus faecalis.