title: A reliable, high-resolution and high-throughput genotyping method for HLA-DRB1.
abstract: Human leukocyte antigen (HLA) DNA typing is an essential part to identify 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.
authors.affiliation: BGI Health Service Co., Ltd, China.
authors.forename: Fan
authors.initials: F
authors.lastname: Yang
authors.affiliation: BGI Health Service Co., Ltd, China.
authors.forename: Fuman
authors.initials: F
authors.lastname: Jiang
authors.affiliation: BGI Health Service Co., Ltd, China.
authors.forename: Xianghua
authors.initials: X
authors.lastname: Chai
authors.affiliation: BGI Health Service Co., Ltd, China.
authors.forename: Yulai
authors.initials: Y
authors.lastname: Guo
authors.affiliation: BGI Health Service Co., Ltd, China.
authors.forename: Zhiheng
authors.initials: Z
authors.lastname: Lan
authors.affiliation: BGI Health Service Co., Ltd, China.
authors.forename: Yuying
authors.initials: Y
authors.lastname: Yuan
authors.affiliation: BGI Health Service Co., Ltd, China.
authors.forename: Changjian
authors.initials: C
authors.lastname: Huang
authors.affiliation: BGI Health Service Co., Ltd, China.
authors.forename: Zhongyi
authors.initials: Z
authors.lastname: Zhu
authors.affiliation: BGI Health Service Co., Ltd, China.
authors.forename: Huiling
authors.initials: H
authors.lastname: Li
authors.affiliation: BGI Health Service Co., Ltd, China.
authors.forename: Bole
authors.initials: B
authors.lastname: Du
authors.affiliation: BGI Health Service Co., Ltd, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Wang
authors.affiliation: BGI Health Service Co., Ltd, China.
authors.forename: Lijun
authors.initials: L
authors.lastname: Zhou
authors.affiliation: BGI Health Service Co., Ltd, China.
authors.forename: Zhonglin
authors.initials: Z
authors.lastname: Chen
authors.affiliation: BGI Health Service Co., Ltd, China.
authors.forename: Ling
authors.initials: L
authors.lastname: Yang
authors.affiliation: BGI Health Service Co., Ltd, China.
authors.forename: Meizhen
authors.initials: M
authors.lastname: Zhou
authors.affiliation: BGI Health Service Co., Ltd, China.
authors.forename: Hao
authors.initials: H
authors.lastname: Zeng
authors.affiliation: BGI Health Service Co., Ltd, China.
authors.forename: Yicou
authors.initials: Y
authors.lastname: Xu
authors.affiliation: BGI Health Service Co., Ltd, China. Electronic address: wangw@genomics.cn.
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
mesh_terms: D000483:Alleles; D017931:DNA Primers; D005091:Exons; D005787:Gene Frequency; D056426:Genetic Loci; D005838:Genotype; D060005:Genotyping Techniques; D059811:HLA-DRB1 Chains / Q000145:classification / Q000235:genetics* / Q000276:immunology; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006650:Histocompatibility Testing; D006801:Humans; D014019:Tissue Donors
chemical_list: D017931:DNA Primers; D059811:HLA-DRB1 Chains
keywords: Ambiguous; Genotyping; High-resolution; Human leukocyte antigen; Next-generation sequencing