Sequence-based typing of HLA: an improved group-specific full-length gene sequencing approach

C.E. Voorter, F. Palusci, M.G. Tilanus

Research output: Chapter in Book/Report/Conference proceedingChapterAcademic

Abstract

Matching for hla at the allele level is crucial for stem cell transplantation. The golden standard approach for allele definition of full gene polymorphism, the so-called high-resolution hla typing, is sequence-based typing (sbt). Although the majority of the polymorphism for class i is located in exons 2 and 3 and for class ii in exon 2, for allele definition it is necessary to unravel the complete coding and intron sequences leading to an ultrahigh hla typing resolution at the allele level, i.e., a full-length gene polymorphism identification.this chapter describes our recently developed sbt method for hla-a, -b, -c, and -dqb1, that is based on full-length hemizygous sanger sequencing of the alleles, separated by group-specific amplification using the low-resolution typing result as reference starting point. Group-specific amplification has already been established for drb. This method enables a cost-efficient, user-friendly sbt approach resulting in a timely unambiguous hla typing to an ultrahigh resolution level with minimal hands-on time.key wordssequence-based typinghuman leukocyte antigenhla class ihla class iifull-length hla gene polymorphismgroup-specific sequencingunambiguous hla typing.
Original languageEnglish
Title of host publicationSequence-based typing of HLA: an improved group-specific full-length gene sequencing approach
PublisherHumana Press
Pages101-114
DOIs
Publication statusPublished - 2014

Publication series

SeriesMethods in Molecular Biology

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