中国食品药品监管Issue(3)：92-103,12.DOI:10.3969/j.issn.1673-5390.2025.03.008
病原靶向高通量测序技术质量评价联合研究
A Joint Study on the Quality Evaluation of Pathogen Targeted Next-Generation Sequencing Technology
摘要
Abstract
Objective:This article aims to conduct a joint study on the quality evaluation of pathogen targeted next-generation sequencing(tNGS)technology to understand the current performance of tNGS reagents in China and to sort out the key factors in quality control,providing a reference for promoting the standardized development of this technology.Methods:Participating institutions used tNGS reagents,developed according to in vitro diagnostic standards,to detect a reference panel containing 30 microorganisms and reported the results.The data were used to analyze the characteristics of tNGS technology selection and its performance.Results:A total of 11 tNGS reagents met the requirements for complete technical documentation and successfully completed detection and data reporting of the reference panel.Among these reagents,9 employed a multiplex amplification approach,while 2 utilized probe capture.The sequencing component included 5 sequencers from 3 different sequencing platforms.The pathogen detection spectrum of each reagent ranged from 30 to 706 targets.The overall performance of the 11 tNGS reagents is as follows:the median positive agreement rate was 88%(67%～100%),the median negative agreement rate was 91%(75%～100%),and the median coefficient of variation was 50%(17%～69%).The detection rates for pathogenic microorganisms at concentration levels of 1×103 CFU/ml and 1×102 CFU/ml(or 1×103 copies/ml and 1×102 copies/ml)were 79%(48%～100%)and 62%(21%～96%),respectively,with a linear correlation coefficient of 0.44(0.18～0.82).Compared with 17 pathogen metagenomic next-generation sequencing(mNGS)reagents,no significant difference was observed in the positive agreement rate of tNGS reagents,although sample cross-contamination had a greater impact on the negative agreement rate of tNGS reagents.Conclusion:This joint study indicates that significant differences exist in the technical choices and performance of various tNGS reagents at this stage,and their overall performance is comparable to that of mNGS reagents as observed in 2019.关键词
病原靶向高通量测序/病原宏基因组高通量测序/质量控制与评价Key words
pathogen targeted next-generation sequencing/pathogen metagenomic next-generation sequencing/quality control and evaluation分类
药学引用本文复制引用
刘东来,王婧,关文达,任珊珊,赵兰青,鲁炳怀,许四宏..病原靶向高通量测序技术质量评价联合研究[J].中国食品药品监管,2025,(3):92-103,12.基金项目
药品监管科学全国重点实验室课题(2024SKLDRS0212) （2024SKLDRS0212）
中国食品药品检定研究院关键技术研究基金(GJJS-2022-2-2) （GJJS-2022-2-2）
