An improved and validated RNA HLA class I SBT approach for obtaining full length coding sequences

K. E. H. Gerritsen, T. I. Olieslagers, M. Groeneweg, C. E. M. Voorter, M. G. J. Tilanus*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

6 Citations (Web of Science)

Abstract

The functional relevance of human leukocyte antigen (HLA) class I allele polymorphism beyond exons 2 and 3 is difficult to address because more than 70% of the HLA class I alleles are defined by exons 2 and 3 sequences only. For routine application on clinical samples we improved and validated the HLA sequence-based typing (SBT) approach based on RNA templates, using either a single locus-specific or two overlapping group-specific polymerase chain reaction (PCR) amplifications, with three forward and three reverse sequencing reactions for full length sequencing. Locus-specific HLA typing with RNA SBT of a reference panel, representing the major antigen groups, showed identical results compared to DNA SBT typing. Alleles encountered with unknown exons in the IMGT/HLA database and three samples, two with Null and one with a Low expressed allele, have been addressed by the group-specific RNA SBT approach to obtain full length coding sequences. This RNA SBT approach has proven its value in our routine full length definition of alleles.
Original languageEnglish
Pages (from-to)450-458
JournalTissue Antigens
Volume84
Issue number5
DOIs
Publication statusPublished - Nov 2014

Keywords

  • full length human leukocyte antigen gene polymorphism
  • HLA class I
  • RNA sequence based typing

Cite this