RESUMO
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.
Assuntos
Éxons , Técnicas de Genotipagem , Cadeias HLA-DRB1/genética , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Alelos , Primers do DNA , Frequência do Gene , Loci Gênicos , Genótipo , Cadeias HLA-DRB1/classificação , Cadeias HLA-DRB1/imunologia , Teste de Histocompatibilidade , Humanos , Doadores de TecidosRESUMO
Twenty-seven novel human leukocyte antigen (HLA) class II alleles in Chinese Marrow Donors are described: 10 HLA-DRB1 alleles and 17 HLA-DQB1 alleles.
