Transfusionsmedizin 2013; 3(4): 196-200
DOI: 10.1055/s-0033-1350835
Praxistipp
Georg Thieme Verlag KG Stuttgart · New York

Next-Generation-Sequencing in der Immungenetik

Next Generation Sequencing in Immunogenetics
C. Gabriel
Blutzentrale Linz, Linz, Österreich
› Institutsangaben
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Publikationsverlauf

Publikationsdatum:
20. November 2013 (online)

Zusammenfassung

Next-Generation-Sequencing ist eine interessante Alternative zu herkömmlichen Verfahren in der hochauflösenden HLA-Typisierung geworden. Insbesondere können mehrdeutige Ergebnisse der Sanger-Sequenzierung aufgelöst werden und eine Vielzahl zusätzlicher Exons getestet werden. Geachtet werden soll aber auf ein robustes System mit möglichst langen Leselängen, die Auswahl der Oliogonukleotide und eine ausreichend hohe Abdeckung der sequenzierten Region.

Abstract

Next generation sequencing is an interesting alternative to existing methods in high resolution HLA-typing. Ambiguous results obtained by Sanger sequencing are resolved and additional exons can be tested. It is important to select a robust system with long reads, to select reliable oligonucleotides and aim at high coverages.

 
  • Literatur

  • 1 Margulies M, Egholm M, Altman WE et al. Genome sequencing in microfabricated high-density picolitre reactors. Nature 2005; 437: 376-380
  • 2 Weir BA, Woo MS, Getz G et al. Characterizing the cancer genome in lung adenocarcinoma. Nature 2007; 450: 893-898
  • 3 Rozera G, Abbate I, Bruselles A et al. Massively parallel pyrosequencing highlights minority variants in the HIV-1 env quasispecies deriving from lymphomonocyte sub-populations. Retrovirology 2009; 6: 15
  • 4 Gabriel C, Danzer M, Hackl C et al. Rapid high-throughput human leukocyte antigen typing by massively parallel pyrosequencing for high-resolution allele identification. Hum Immunol 2009; 70: 960-964
  • 5 Stabentheiner S, Danzer M, Niklas N et al. Overcoming methodical limits of standard RHD genotyping by next-generation sequencing. Vox Sang 2011; 100: 381-388
  • 6 Petersdorf EW. Optimal HLA matching in hematopoietic cell transplantation. Curr Opin Immunol 2008; 20: 588-593
  • 7 Adams SD, Barracchini KC, Chen D et al. Ambiguous allele combinations in HLA class I and class II sequence-based typing: when precise nucleotide sequencing leads to imprecise allele identification. J Transl Med 2004; 2: 30
  • 8 Metzker ML. Sequencing technologies – the next generation. Nat Rev Genet 2010; 11: 31-46
  • 9 Niklas N, Pröll J, Danzer M et al. Routine performance and errors of 454 HLA exon sequencing in diagnostics. BMC Bioinformatics 2013; 14: 176
  • 10 Lank SM, Wiseman RW, Dudley DM et al. A novel single cDNA amplicon pyrosequencing method for high-throughput, cost-effective sequence-based HLA class I genotyping. Hum Immunol 2010; 71: 1011-1017