Full-Length Characterization of Novel HLA-DRB1 Alleles for Reference Database Submission.
Methods Mol Biol
; 2809: 145-156, 2024.
Article
em En
| MEDLINE
| ID: mdl-38907896
ABSTRACT
The prerequisite for successful HLA genotyping is the integrity of the large allele reference database IPD-IMGT/HLA. Consequently, it is in the laboratories' best interest that the data quality of submitted novel sequences is high. However, due to its long and variable length, the gene HLA-DRB1 presents the biggest challenge and as of today only 16% of the HLA-DRB1 alleles in the database are characterized in full length. To improve this situation, we developed a protocol for long-range PCR amplification of targeted HLA-DRB1 alleles. By subsequently combining both long-read and short-read sequencing technologies, our protocol ensures phased and error-corrected sequences of reference grade quality. This dual redundant reference sequencing (DR2S) approach is of particular importance for correctly resolving the challenging repeat regions of DRB1 intron 1. Until today, we used this protocol to characterize and submit 384 full-length HLA-DRB1 sequences to IPD-IMGT/HLA.
Palavras-chave
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Bases de Dados Genéticas
/
Alelos
/
Cadeias HLA-DRB1
Limite:
Humans
Idioma:
En
Ano de publicação:
2024
Tipo de documento:
Article