杂交探针捕获与扩增子文库构建法对HLA基因分型分辨水平的影响  

Influence of hybridization probe capture and amplicon library construction methods on HLA genotyping resolution level

作　　者：袁晓妮[1] 张腾腾[1] 李杨[1] 姜雪 杨天杰[1] 鲍晓晶[1] 何军[1] YUAN Xiaoni;ZHANG Tengteng;LI Yang;JIANG Xue;YANG Tianjie;BAO Xiaojing;HE Jun(The First Affiliated Hospital of Soochow University,Jiangsu Provincial Institute of Hematology,Suzhou 215006,China)

机构地区：[1]苏州大学附属第一医院江苏省血液研究所HLA配型实验室,江苏苏州215006

出　　处：《中国输血杂志》2025年第3期303-308,321,共7页Chinese Journal of Blood Transfusion

基　　金：国家自然科学基金(82070180);江苏高校血液学协同创新中心(SX21100121)。

摘　　要：目的 比较高通量测序(next generation sequencing, NGS)杂交探针捕获和扩增子文库构建方法,分析HLA基因分型分辨水平的影响因素及临床应用中的前景。方法 选择207例已知分型结果临床标本和能力验证计划标本,确认HLA基因分型结果符合率、等位基因覆盖度及分型数据分析指标,比较2种文库构建方法对HLA基因分型分辨水平的影响。结果 207例标本杂交探针捕获法结果与能力验证反馈结果或已知结果符合率均为100%。其中91例标本与原扩增子法比较,杂交探针捕获法可区分13例标本DRB1、DPB1不能确定的等位基因;解决6例标本存在DRB1、DPA1、DQB1位点等位基因不平衡;明确3例患者标本存在DQA1、DQB1位点等位基因漏检。结论 杂交探针捕获与扩增子法均满足HLA基因分型临床检测的要求,本文为其临床推广应用提供了实验方法和依据。Objective To compare next generation sequencing(NGS)library construction technology between probe hybridization capture and amplicon methods,and analyze the influencing factors of HLA genotyping resolution level and its prospects in clinical applications.Methods A total of 207 clinical samples with known typing results and samples from the proficiency testing plan were selected.The conformity rate of HLA genotyping results,allele coverage and typing data analy-sis indicators were confirmed,and the effects of two library construction methods on the level of HLA genotyping discrimina-tion were compared.Results The concordance rate of 207 samples with the feedback results of PT or prior well-character-ized HLA genotypes was 100%.Among them,91 samples were captured using hybridization probe capture method.Com-pared with the original amplicon method,the hybridization probe capture method can distinguish the alleles of DRB1 and DPB1 that cannot be determined in 13 samples.The allelic imbalance of DRB1,DPA1,and DQB1 loci in 6 samples was re-solved.Three samples were found to have missed detection of alleles at the DQA1 and DQB1 loci.Conclusion The per-formance indicators of hybridization probe capture and amplicon performance confirmation meet the requirements of clinical detection of HLA genotyping,which provides an experimental method and basis for clinical application.

关 键 词：HLA 高通量测序 等位基因 临床应用 

分 类 号：R446[医药卫生—诊断学]