同源重组修复缺陷检测国家参考品  

National reference materials for homologous recombination deficiency detection

作　　者：张咪 张文新 孙楠[1] 于婷[1] 胡泽斌[1] 黄杰[1] 曲守方[1] ZHANG Mi;ZHANG Wenxin;SUN Nan;YU Ting;HU Zebin;HUANG Jie;QU Shoufang(National Institutes for Food and Drug Control,Beijing,China,100050)

机构地区：[1]中国食品药品检定研究院,北京100050

出　　处：《分子诊断与治疗杂志》2024年第11期2112-2115,共4页Journal of Molecular Diagnostics and Therapy

基　　金：中国食品药品检定研究院关键技术研究基金(GJJS⁃2022⁃2⁃1)。

摘　　要：目的制备同源重组修复缺陷检测(HRD)国家参考品,并建立其标准集,用于HRD体外诊断试剂产品的性能评价。方法采用肿瘤组织永生化的细胞和配对白细胞永生化的细胞,制备不同DNA比例(50%,40%,30%,20%,10%)的HRD国家参考品。将其基因组DNA打断,进行全外显子测序(WES),将获得的测序数据进行分析。首先合并拷贝数变异(CNV)检测大于10 Mb区域,构建成区间树。然后将区间树拆分成最小节点,统计各节点检出频次及节点中各类型检出频次,建立等位基因特异性拷贝数(ASCNV)标准集。使用scarHRD软件建立LOH、LST及TAI标准集。结果ASCNV标准集包括节点检出频率≥50%的节点,且节点合并后长度≥3 Mb。建立了国家参考品的LOH、LST及TAI三个指标的标准集。结论HRD国家参考品和标准集,可以为同源重组修复缺陷检测试剂盒的性能评价提供技术支撑。Objective To prepare national reference materials for homologous recombination repair defect detection(HRD)and establish its standard set for performance evaluation of HRD in vitro diagnostic reagent products.Methods Immortalized cells from tumor tissue and paired white blood cells are used to prepare HRD national reference materials with different DNA ratios(50%,40%,30%,20%,10%).The genomic DNA is then broken down,and whole exome sequencing(WES)is performed to obtain and analyze the sequencing data.The detection regions of copy number variation larger than 10Mb are merged,and an interval tree is con⁃structed.This interval tree is then split into minimum nodes and the detection frequency of each node and the detection frequency of each type in the node are counted to establish an ASCNV standard set.The scarHRD software is used to acquire the standard set for LOH,LST,and TAI indicators.Results The ASCNV standard set includes nodes with a type of detection frequency of≥50%,and a merged length of≥3 Mb.The standard set is established for LOH,LST,and TAI.Conclusion The HRD national reference and standard set provides technical support for evaluating the performance of kits for detecting defects in homologous recombination repair.

关 键 词：同源重组修复缺陷 聚腺苷二磷酸核糖聚合酶抑制剂 生物标志物 下一代测序 全外显子测序 标准集 单核苷酸多态性 

分 类 号：R927.1[医药卫生—药学]