中国组织工程研究

中国组织工程研究 ›› 2021, Vol. 25 ›› Issue (25): 4009-4012.doi: 10.12307/2021.012

• 干细胞基础实验 basic experiments of stem cells • 上一篇    下一篇

第2代测序技术鉴定人类白细胞抗原等位基因A*24:353及基因频率分析

何柳媚，陈  浩，钟艳平，全湛柔，邹红岩   

  1. 深圳市血液中心输血医学研究所，广东省深圳市  518020
  • 收稿日期:2020-06-05 修回日期:2020-06-06 接受日期:2020-07-14 出版日期:2021-09-08 发布日期:2021-03-26
  • 通讯作者: 邹红岩，主任技师，深圳市血液中心输血医学研究所，广东省深圳市 518020
  • 作者简介:何柳媚，女，1980年生，广东省梅州市人，汉族，2005年暨南大学毕业，硕士，副主任技师，主要从事人白细胞抗原组织配型高分辨确认工作。

Next-generation sequencing of identifying the human leukocyte antigen-A*24:353 and its gene frequency analysis

He Liumei, Chen Hao, Zhong Yanping, Quan Zhanrou, Zou Hongyan   

  1. Institute of Transfusion Medicine of Shenzhen Blood Center, Shenzhen 518020, Guangdong Province, China
  • Received:2020-06-05 Revised:2020-06-06 Accepted:2020-07-14 Online:2021-09-08 Published:2021-03-26
  • Contact: Zou Hongyan, Chief technician, Institute of Transfusion Medicine of Shenzhen Blood Center, Shenzhen 518020, Guangdong Province, China
  • About author:He Liumei, Master, Associate chief technician, Institute of Transfusion Medicine of Shenzhen Blood Center, Shenzhen 518020, Guangdong Province, China

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摘要：

文题释义：
人类白细胞抗原(human leukocyte antigen，HLA)：是由人类白细胞基因复合体所编码的产物，位于第6号染色体短臂上。临床干细胞移植前配型需检测供受者HLA-A、B、C、DRB1、DQB1五个基因位点相合的程度，供受者的HLA配型达到一定相合要求才能进行造血干细胞移植。
第2代测序技术(next generation sequencing，NGS)：又称“下一代测序技术”，是一种以“边合成边测序”为核心思想的高通量测序技术，能够在短时间内同时对上百亿碱基进行全长序列测定。

背景：人类白细胞抗原(human leukocyte antigen，HLA)高分辨确认工作是造血干细胞移植前必不可少的步骤之一，随着工作的不断深入开展及被检测人群的持续增加，人类白细胞抗原新等位基因不断涌现。有些新基因由于发现较晚，相关报道较少，导致基因频率不能准确计算，在缺乏相应数据的情况下，容易造成分型结果的误判，导致配型不相合结果，影响移植配型成功率。
目的：对HLA一个等位基因HLA-A*24:353进行检测确认，并对其出现频率进行分析。
方法：应用聚合酶链反应-直接测序分型方法对2019年9月至2020年5月间543例临床移植配型供受者进行HLA-A、-B、-C、-DRB1、-DQB1五个位点检测，再用第2代测序技术对HLA-A*24:02/24:353模棱两可结果进行全长序列测定，得到HLA高分辨分型结果，并计算HLA-A*24:353基因出现频率。
结果与结论：检出HLA-A*24:02/24:353模棱两可结果146例，采用第2代测序技术进一步鉴定后，检出HLA-A*24:353基因5例，占A*24组的比例为3.42%，在543例检测人群中出现比例为0.92%。结果提示HLA-A*24:353在中国可能并不罕见，进行临床移植配型供受者高分辨分型时，当出现HLA-A*24:02/24:353模棱两可结果时，需做进一步确认，以增加分型结果的准确性，提高移植配型的成功率。
https://orcid.org/0000-0001-5840-011X(何柳媚) 

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

关键词: 造血干细胞, 移植, 人类白细胞抗原, 等位基因, 频率；移植配型, 血液

Abstract:

BACKGROUND: High resolution validation of human leukocyte antigen (HLA) is one of the essential step before hematopoietic stem cell transplantation. With the continuous development of the work and the number of people tested continues to increase, human leukocyte antigen new alleles are constantly emerging. Due to the late discovery of some new genes and the lack of relevant reports, the gene frequency cannot be accurately calculated. In the absence of relevant data, it is easy to misjudge the typing results. The result of mismatch affects the success rate of transplantation. 

OBJECTIVE: An allele of HLA-A*24:353 was detected and confirmed in HLA, and its occurrence frequency was analyzed.
METHODS:  Polymerase chain reaction-sequencing-based typing method was used to detect five loci of HLA- A, B, C, DRB1, and DQB1 in 543 clinical transplantation matching recipients from September 2019 to May 2020. Next-generation sequencing was used to determine the full-length sequence of HLA-A*24:02/24:353 ambiguous results to obtain high resolution typing results of HLA and calculate the occurrence frequency of HLA-a *24:353 gene.
RESULTS AND CONCLUSION: The ambiguous results of HLA-A*24:02/24:353 were detected in 146 cases. After further confirmatory tests using next-generation sequencing, the results of HLA-A*24:353 gene were detected in 5 cases (3.42%) in A*24 group and 0.92% in 543 cases. The results suggest that HLA-A*24:353 may not be rare allele in China. When HLA-A*24:02/24:353 ambiguous results appear in clinical transplantation matching recipients for high resolution typing, further confirmation is needed to increase the accuracy of typing results and improve the success rate of transplantation matching.

Key words: hematopoietic stem cells, transplantation, human leukocyte antigen, allele, frequency, transplantation matching, blood

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