病原菌多重核酸检测试剂盒分析性能质量评价研究

Quality evaluation study of the analytical performance of multiplex nucleic acid detection kits for pathogenic microorganisms

目的了解我国病原菌多重核酸检测试剂盒分析性能的现状,特别是检出限性能的基本情况,为相关研发企业和医疗机构使用者提供参考。方法根据体外诊断试剂行业标准《细菌和真菌感染多重核酸检测试剂盒(标准号YY∕T 1725-2020)》,使用“34种细菌和真菌感染多重核酸检测试剂国家参考品(批号370026-201801)”对10种不同检测技术原理的病原菌多重核酸检测试剂盒的分析性能进行评估,包括阳性符合率、阴性符合率、重复性及检出限等,并按照国家参考品的质量标准对评价结果进行统计分析。结果10款试剂盒中包括4款呼吸道感染试剂盒、3款中枢神经系统感染试剂盒及3款血流感染试剂盒,检测范围或病原谱覆盖8种革兰阳性细菌、14种革兰阴性细菌及10种真菌和非典型病原体。各试剂盒的阳性符合率、阴性符合率、重复性及检出限等分析性能均符合国家参考品的质量标准。呼吸道、中枢神经系统及血流感染试剂盒病原菌谱及检出限性能差异较大,检出限的中位数分别为5×10^(3)CFU/ml、1×10^(3)CFU/ml及1×10^(6)CFU/ml;各试剂盒病原菌谱中分布和出现频率较高的6种病原菌检出限中位值分别为5×10^(2)CFU/ml(肺炎链球菌)、3×10^(3)CFU/ml(金黄色葡萄球菌的)、1×10^(3)CFU/ml(流感嗜血杆菌)、5×10^(4)CFU/ml(铜绿假单胞菌)、3×10^(6)CFU/ml(鲍曼不动杆菌)及5×10^(4)CFU/ml(大肠埃希菌)。结论我国已经进入体外诊断试剂注册阶段的病原菌多重核酸检测试剂盒均具有较好的准确性、特异性及重复性。同时,研发企业应注意结合临床具体的需求,设计合理的试剂盒病原菌谱,并持续优化检出限性能。

Objective Understand the current update in the analytical performance of multiplex nucleic acid detection kits for pathogenic microorganisms in China,especially the basic situation of detection limit,and provide a reference to users of related enterprises and medical institutions.Methods The analytical performance of the multiplex nucleic acid detection kits of 10 different detection technology principles has been evaluated,according to the in vitro diagnostic reagent industry standard of“Multiplex Nucleic Acid Detection Kit for Infection of Microorganisms(YY/T 1725-2020)”and“National Reference Panel for Multiplex Nucleic Acid Assay for Identification of Infection of 34 Microorganisms(370026-201801)”.It includes positive coincidence rate,negative coincidence rate,repeatability and limit of detection(LOD).Besides,the evaluation results were summarized and analyzed according to the quality standard of the national reference panel.Results The 10 kits included 4 respiratory infection kits,3 central nervous system infection kits,and 3 bloodstream infection kits,and the detection range or pathogenic microorganism spectrum covered 8 Gram-positive bacteria,14 Gram-negative bacteria,and 10 fungi and atypical pathogens.The positive coincidence rate,negative coincidence rate,repeatability,and LOD of each kit were all in line with the quality standard of the national reference panel.Furthermore,the 6 pathogens with high distribution and frequency in each kit were Streptococcus pneumoniae(median detection limit 5×10^(2) CFU/ml),Staphylococcus aureus(median detection limit 3×10^(3) CFU/ml),Haemophilus influenzae(median detection limit 1×10^(3) CFU/ml),Pseudomonas aeruginosa(median detection limit 5×10^(4) CFU/ml),Acinetobacter baumannii(median detection limit 3×10^(6) CFU/ml)and Escherichia coli(median detection limit 5×10^(4) CFU/ml).Conclusions The multiplex nucleic acid detection kits for pathogenic microorganisms have high accuracy,specificity,and repeatability,which have entered the registration stage of in vitro diagnostics in China.Meanwhile,the relevant research and development enterprises should pay attention to combining clinical needs and design a reasonable kit pathogen spectrum and optimize LOD performance.