HLA-DQB1*03:90N碱基缺失的序列分析及确认  

Analysis and verification of a HLA-DQB1*03:90N allele with a single base deletion

作　　者：全湛柔 邹红岩[1] 陈浩 钟艳平[1] 周丹[1] 邓志辉[1] 洪文旭[1] Quan Zhanrou;Zou Hongyan;Chen Hao;Zhong Yanping;Zhou Dan;Deng Zhihui;Hong Wenxu(Institute of Transfusion Medicine,Shenzhen Blood Center,Shenzhen,Guangdong 518020,China)

机构地区：[1]深圳市血液中心输血医学研究所,518020

出　　处：《中华医学遗传学杂志》2020年第6期681-684,共4页Chinese Journal of Medical Genetics

基　　金：深圳市卫生计生系统科研项目(201506076);深圳市医疗卫生三名工程项目(SZSM201811092)。

摘　　要：目的确认人类白细胞抗原罕见等位基因HLA-DQB1*03:90N的序列,探讨检测方法,以提高HLA分型的准确性。方法采用PCR-序列特异性寡核苷酸探针(sequence-specific oligonudeotide probe,SSOP)对2018年中国造血干细胞捐献者资料库深圳分库登记的2265例造血干细胞捐献者进行HLA分型检测,针对1例HLA-DQB1罕见等位基因进一步选择聚合酶链反应-直接测序分型(sequence-based tying,SBT)技术和基于Ion Torrent S5平台的二代测序(next generation sequencing,NGS)分析技术进行确认。结果HLA-DQB1位点SSOP分型结果显示为罕见等位基因,经SBT复核发现序列在第2外显子疑似插入或缺失,最后通过NGS确认结果为DQB1*03:90N,DQB1*06:01。结论经SSOP法检测得到的罕见等位基因不能轻易排除,应借助多种方法复核确认,保证HLA基因分型结果的准确性。罕见等位基因或新等位基因序列存在碱基缺失,采用二代测序技术可能得到更准确的结果。Objective To verify a HLA-DQB1*03:90N allele and method to improve the accuracy of HLA typing.Methods A total of 2265 hematopoietic stem cell donors from Shenzhen Branch of China Marrow Donor Program in 2018 were initially detected by a PCR sequence-specific oligonucleotide probe(SSOP)method.Among these,a rare HLA-DQB1 allele was identified by sequence-based tying(SBT)and Ion Torrent S5 next generation sequencing(NGS).Results The SSOP typing result suggested the HLA-DQB1 to be a rare allele,while an insertion and a deletion was suspected in its exon 2 by SBT,which were confirmed by NGS as DQB1*03:90N and DQB1*06:01,respectively.Conclusion Rare alleles suspected by the SSOP method should be verified by other methods to ensure the accuracy of HLA genotyping.Rare alleles formed by deletions can be detected by NGS with accuracy.

关 键 词：HLA-DQB1 碱基缺失 聚合酶链反应-序列特异性寡核苷酸探针 聚合酶链反应-直接测序分型 二代测序 

分 类 号：R457.11[医药卫生—治疗学] R440[医药卫生—临床医学]