Abstract
DNA sequencing is a powerful technique for identifying allelic variation within the human leukocyte antigen (HLA) genes. Sequencing is usually focused on the most polymorphic exons of the class I (HLA-A, -B, -C) and class II (HLA-DR, -DQ, and -DP) genes. These exons encode the antigen recognition site, the region of the HLA molecule that binds peptides and interacts with the T cell receptor for antigen and natural killer cell immunoglobulin-like receptors (KIR). Sanger sequencing of amplified DNA from each HLA gene from a preparation containing one or two alleles yields a sequence that is used to identify the alleles by comparison with a reference database.
Ana Lazaro and Bin Tu have contributed equally to first authorship.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Posch PE, Hurley CK (2011) Histocompatibility: HLA and other systems. In: Porwit A, McCullough J, Erber WN (eds) Blood and bone marrow pathology, 2nd edn. Elsevier, Edinburgh, pp 641–676
Robinson J, Waller MJ, Fail SC, McWilliam H, Lopez R, Parham P et al (2009) The IMGT/HLA database. Nucleic Acids Res 37(Database issue):D1013–D1017
Sanger F, Nicklen S, Coulson A (1977) DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
Bray RA, Hurley CK, Kamani NR, Woolfrey A, Muller C, Spellman S et al (2008) National marrow donor program HLA matching guidelines for unrelated adult donor hematopoietic cell transplants. Biol Blood Marrow Transplant 14(9 Suppl):45–53
Lee SJ, Klein J, Haagenson M, Baxter-Lowe LA, Confer DL, Eapen M et al (2007) High-resolution donor-recipient HLA matching contributes to the success of unrelated donor marrow transplantation. Blood 110(13):4576–4583
Barker JN, Scaradavou A, Stevens CE (2010) Combined effect of total nucleated cell dose and HLA match on transplantation outcome in 1061 cord blood recipients with hematologic malignancies. Blood 115(9):1843–1849
Kamani N, Spellman S, Hurley CK, Barker JN, Smith FO, Oudshoorn M et al (2008) State of the art review: HLA matching and outcome of unrelated donor umbilical cord blood transplants. Biol Blood Marrow Transplant 14(1):1–6
Spellman S, Bray R, Rosen-Bronson S, Haagenson M, Klein J, Flesch S et al (2010) The detection of donor-directed, HLA-specific alloantibodies in recipients of unrelated hematopoietic cell transplantation is predictive of graft failure. Blood 115(13):2704–2708
Kotsch K, Wehling J, Blasczyk R (1999) Sequencing of HLA class II genes based on the conserved diversity of the non-coding regions: sequencing based typing of HLA-DRB genes. Tissue Antigens 53:486–497
Dunn PP, Day S, Williams S, Bendukidze N (2005) HLA-DQB1 sequencing-based typing using newly identified conserved nucleotide sequences in introns 1 and 2. Tissue Antigens 66(2):99–106
Cao K, Chopek M, Fernandez-Vina MA (1999) High and intermediate resolution DNA typing systems for Class I HLA-A, -B, -C genes by hybridization with sequence specific oligonucleotide probes (SSOP). Rev Immunogenet 1:177–208
Cereb N, Maye P, Lee S, Kong Y, Yang SY (1995) Locus-specific amplification of HLA class I genes from genomic DNA: locus-specific sequences in the first and third introns of HLA-A, -B, and -C alleles. Tissue Antigens 45(1):1–11
Cereb N, Yang SY (1997) Dimorphic primers derived from intron 1 for use in the molecular typing of HLA-B alleles. Tissue Antigens 50(1):74–76
Bochtler W, Maiers M, Oudshoorn M, Marsh SG, Raffoux C, Mueller C et al (2007) World Marrow Donor Association guidelines for use of HLA nomenclature and its validation in the data exchange among hematopoietic stem cell donor registries and cord blood banks. Bone Marrow Transplant 39(12):737–741
Summers CW, Hampson VJ, Taylor GM (1993) HLA class I non-coding nucleotide sequences, 1992. Eur J Immunogenet 20(3):201–240
Blasczyk R, Kotsch K, Wehling J (1998) The nature of polymorphism of the HLA-DRB intron sequences is lineage specific. Tissue Antigens 52:19–26
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2013 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Lazaro, A. et al. (2013). Human Leukocyte Antigen (HLA) Typing by DNA Sequencing. In: Zachary, A., Leffell, M. (eds) Transplantation Immunology. Methods in Molecular Biology, vol 1034. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-493-7_9
Download citation
DOI: https://doi.org/10.1007/978-1-62703-493-7_9
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-492-0
Online ISBN: 978-1-62703-493-7
eBook Packages: Springer Protocols