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

作者：Yang, Fan; Jiang, Fuman; Chai, Xianghua; Guo, Yulai; Lan, Zhiheng; Yuan, Yuying; Huang, Changjian; Zhu, Zhongyi; Li, Huiling; Du, Bole; Wang, Jun; Zhou, Lijun; Chen, Zhonglin; Yang, Ling; Zhou, Meizhen; Zeng, Hao; Xu, Yicou; Wang, Wei^*

来源：Human Immunology, 2015, 76(6): 397-401.

DOI：10.1016/j.humimm.2015.04.003

• Ambiguous
• Human leukocyte antigen
• High-resolution
• Genotyping
• Next-generation sequencing

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.

• 出版日期2015-6