Strategies and technical challenges in allele level Class II typing in 2578 bone marrow transplantation donor–recipient pairs

Summary Human leukocyte antigen typing of 2578 donor–recipient pairs whose transplantation was facilitated by the National Marrow Donor Program allowed for an in-depth analysis of the accuracy of high-volume allele level testing data. The methods employed provided allele level typing at DRB1/3/5, DQ...

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Bibliographic Details
Published in:Human immunology 2008-04, Vol.69 (4), p.227-234
Main Authors: Williams, T.M, Winden, T, Setterholm, M, Vierra-Green, C.A, Spellman, S, Flesch, S, Awdeh, Z, Baxter-Lowe, L.A, Begovich, A.B, Fernandez-Vina, M, Hegland, J, Hurley, C.K, Johnson, D, Noreen, H, Salazar, M, Schmeckpeper, B, Yunis, E.J
Format: Article
Language:English
Subjects:
Alleles
Allergy and Immunology
Bone Marrow Transplantation
Histocompatibility antigens class II
Histocompatibility Testing - methods
HLA antigens
HLA-D Antigens - genetics
Humans
Nucleic Acid Hybridization
Oligonucleotide Probes
Polymerase Chain Reaction
Polymorphism, Genetic
Polymorphism, Restriction Fragment Length
Reproducibility of Results
Retrospective Studies
Sequence Analysis, DNA - methods
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Summary:Summary Human leukocyte antigen typing of 2578 donor–recipient pairs whose transplantation was facilitated by the National Marrow Donor Program allowed for an in-depth analysis of the accuracy of high-volume allele level testing data. The methods employed provided allele level typing at DRB1/3/5, DQA1, DQB1, DPA1, and DPB1 using sequence-specific oligonucleotide probe hybridization (SSOPH), polymerase chain reaction (PCR) restriction fragment length polymorphism analysis, sequence specific PCR, and direct sequence-based typing (SBT). Each typing was independently tested by two laboratories in Phase 1, and in subsequent phases targeted samples were typed in duplicate by SBT to monitor typing quality. Comparison with prior transplant center typing was also evaluated. SSOPH detected discrepancies ranged from 0.6% at DPB1 to 5.1% at DQB1 in Phase 1. The majority of discrepancies, 62%, resulted from human error such as sample handling, result interpretation, or clerical errors. Alleles that are frequently discrepant have been identified in this predominantly white population.
ISSN:0198-8859
1879-1166
DOI:10.1016/j.humimm.2008.03.004