A reliable, high-resolution and high-throughput genotyping method for HLA-DRB1

被引：1

作者：

Yang, Fan ^{[1
]}

Jiang, Fuman ^{[1
]}

Chai, Xianghua ^{[1
]}

Guo, Yulai ^{[1
]}

Lan, Zhiheng ^{[1
]}

Yuan, Yuying ^{[1
]}

Huang, Changjian ^{[1
]}

Zhu, Zhongyi ^{[1
]}

Li, Huiling ^{[1
]}

Du, Bole ^{[1
]}

Wang, Jun ^{[1
]}

Zhou, Lijun ^{[1
]}

Chen, Zhonglin ^{[1
]}

Yang, Ling ^{[1
]}

Zhou, Meizhen ^{[1
]}

Zeng, Hao ^{[1
]}

Xu, Yicou ^{[1
]}

Wang, Wei ^{[1
]}

机构：

[1] BGI Hlth Serv Co Ltd, Shenzhen, Peoples R China

来源：

HUMAN IMMUNOLOGY | 2015年 / 76卷 / 06期

关键词：

Ambiguous; Human leukocyte antigen; High-resolution; Genotyping; Next-generation sequencing; HLA-B; PCR; ACCURATE; LEVEL; DNA;

D O I：

10.1016/j.humimm.2015.04.003

中图分类号：

R392 [医学免疫学]; Q939.91 [免疫学];

学科分类号：

100102 ;

摘要：

Human leukocyte antigen (HLA) DNA typing is an essential part to identity a donor who may be a good match for the patients who need haematopoietic stem cells from bone marrow. Thus, fetching quickly high-resolution genotype is very important at present. However, current HLA DNA typing methods especially for HLA-DRB1 generally yielded ambiguous typing results because of high degree of heterozygosity on exome region and primer pairs design limitations, which generally amplified only exon2 of HLA-DRB1 and the position of its primer pairs is on exome region. To solve this problem, we developed a reliable, high-resolution and high-throughput (RHH) sequence based typing approach for HLA-DRB1 through massively parallel paired-end sequencing. Several primer pairs were designed to amplify three exon regions, which are part of exon1, the whole region of exon2 and exon3 of HLA-DRB1, to genotyping for HLA-DRB1 by synthesis. 94 samples were included to analyze and the highly successful genotyping ratio (98.94%) and no ambiguous genotyping result demonstrated the accurate performance of our method for HLA-DRB1 genotyping. (C) 2015 American Society for Histocompatibility and Immunogenetics. Published by Elsevier Inc. All rights reserved.

引用

页码：397 / 401

页数：5

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