【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2 076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。更多还原

【Abstract】 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 2 076 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-B* 46:18, 54:06 or HLA-B*46:01, 54:XX(including a base mutation), and nanopore sequencing confirmed the typing as HLA-B*46:01, 54:XX(including a base mutation). Compared with HLA-B*54:01:01:01, the HLA-B*54: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-B*54: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-B*54:01:11 by the World Health Organization Nomenclature Committee for Factors of the HLA System.更多还原