国产试剂盒在二代测序平台的性能验证实验  

Performance verification experiments of domestic reagent kits on next-generation sequencing platform

作　　者：贺晓花 乔一芸 魏波 王国洪 牟永平 HE Xiaohua;QIAO Yiyun;WEI Bo;WANG Guohong;MOU Yongping(Department of Laboratory Medicine,Affiliated Tumour Hospital of Inner Mongolia Medical University,Inner Mongolia,Hohhot 010020,China)

机构地区：[1]内蒙古医科大学附属肿瘤医院检验科,内蒙古呼和浩特010020

出　　处：《中国医药科学》2025年第7期166-170,共5页China Medicine And Pharmacy

基　　金：内蒙古自治区卫生健康委科学基金资助项目(202203158);北京大学肿瘤医院内蒙古医院“青苗”人才计划(QM202332)。

摘　　要：目的利用二代测序(NGS)平台对多基因突变国产测序试剂盒进行性能验证实验,评估其在临床检测中的准确性、精密性和一致性,为医疗机构开展测序项目提供技术参考。方法使用标准参考品和临床样本对5家公司的国产试剂盒进行检测,并比较检测过程和结果。结果5种测序试剂盒的性能分析表明,准确性检测即对阳性和阴性标准品检出均为100%。最低检测等位基因突变频率百分比参考值含1%~7%梯度突变频率和5个拷贝数的扩增变异,结果显示相关系数(r)在0.75~0.93(P<0.001)。精密度检测,每个试剂盒检测结果的变异系数(CV)均值小于11%,Kruskal-Wallis检验P=0.965。结论国产试剂盒在NGS平台验证实验结果表明,在准确度、灵敏度和精密度方面均符合检测标准。5个临床样本的检测结果与预期结果一致,为临床检测工作提供了技术指标依据。Objective To conduct performance verification experiments of domestic sequencing kits for multigene mutations using the next-generation sequencing(NGS)platform,to evaluate their accuracy,precision,and consistency in clinical testing,and to provide technical references for medical institutions to carry out sequencing projects.Methods Standard reference materials and clinical samples were used to test the domestic reagent kits of 5 companies,and the testing process and results were compared.Results The performance analysis of the 5 sequencing kits showed that the accuracy detection was 100%for both positive and negative standard samples.The reference value for the lowest detected allele mutation frequency percentage included 1%-7%gradient mutation frequency and 5 copy number amplification variations.The results showed a correlation coefficient(r)between 0.75-0.93(P<0.001).Precision testing showed that the mean coefficient of variation(CV)of each test kit was less than 11%,with a Kruskal-Wallis test P=0.965.Conclusion The verification experiment results of the domestic reagent kits on the NGS platform show that it meets the detection standards in terms of accuracy,sensitivity,and precision.The test results of 5 clinical samples are consistent with the expected results,providing technical indices for clinical testing work.

关 键 词：二代测序 性能验证 肿瘤 临床评估 

分 类 号：R446[医药卫生—诊断学]