应用三代测序技术鉴定NGS方法检出的新等位基因HLA-B*54:01:11  

Identification of the Novel Allele HLA-B54:01:11 Detected by NGS Using the Third Generation Sequencing Technology

作　　者：陈男英[1] 何亿镇 皮雯雯 李奇 董丽娜[1] 章伟[1] CHEN Nan-Ying;HE Yi-Zheng;PI Wen-Wen;LI Qi;DONG Li-Na;ZHANG Wei(Institute of Transfusion Medicine,Blood Center of Zhejiang Province,Hangzhou 310052,Zhejiang Province,China)

机构地区：[1]浙江省血液中心输血医学研究所,浙江杭州310052

出　　处：《中国实验血液学杂志》2025年第2期565-568,共4页Journal of Experimental Hematology

基　　金：浙江省自然科学基金(LTGY24H080007);浙江省医药卫生科学研究基金(2023KY658);浙江省医药卫生科学研究基金(2023KY087)。

摘　　要：目的:区分人类白细胞抗原(HLA)模棱两可的基因分型结果,鉴定HLA-B新等位基因,并分析其核苷酸序列。方法:利用基于Ion Torrent S5平台的二代测序技术(NGS)对2022年2076例浙江省脐带血库样本进行HLA入库分型检测,发现1例含碱基突变的模棱两可组合样本,选择基于纳米孔技术平台的三代测序技术(TGS)对其进行鉴定。结果:HLA-B位点经NGS检测分型结果显示为HLA-B*46:18,54:06/46:01,54:XX(含碱基突变)组合,经纳米孔测序鉴定结果为HLA-B*46:01,54:XX(含碱基突变)。HLA-B*54:XX与同源性最高的HLA-B*54:01:01:01相比,第6外显子1014位碱基T>C,并未引起任何氨基酸的改变。新等位基因序列已递交给GenBank数据库(OP853532)。结论:应用纳米孔测序技术区分了1例NGS方法产生的模棱两可结果,并成功鉴定了HLA-B新等位基因。该新等位基因被世界卫生组织HLA因子命名委员会正式命名为HLA-B*54:01:11。Objective:To distinguish the ambiguous genotyping results of human leukocyte antigen(HLA),identify a novel HLA-B allele and analyze the nucleotide sequence.Methods:A total of 2076 umbilical core blood samples from the Zhejiang Cord Blood Bank in 2022 were detected using the next generation sequencing technology(NGS)based on the Ion Torrent S5 platform.Among these a rare HLA-B allele with ambiguous combination result containing a base mutation was identified,and was further confimed by the third-generation sequencing(TGS)based on the nanopore technology.Results:The NGS typing result of HLA-B locus showed HLA-B46:18,54:06 or HLA-B46:01,54:XX(including a base mutation),and nanopore sequencing confirmed the typing as HLA-B46:01,54:XX(including a base mutation).Compared with HLA-B54:01:01:01,the HLA-B54:XX allele showed one single nucleotide substitution at position 1014 T>C in exon 6,with no amino acid change.The nucleotide sequence of the novel HLA-B54:XX has been submitted to the GenBank nucleotide sequence database and the accession number OP853532 was assigned.Conclusion:A ambiguous genotyping of the HLA-B Locus detected by NGS was distinguished by nanopore sequencing and a new HLA-B allele was successfully identified,which was officially named as HLA-B54:01:11 by the World Health Organization Nomenclature Committee for Factors of the HLA System.

关 键 词：HLA-B 新等位基因 二代测序 纳米孔测序 碱基突变 

分 类 号：R440[医药卫生—诊断学] R394[医药卫生—临床医学]