title: Strategies and technical challenges in allele level Class II typing in 2578 bone marrow transplantation donor-recipient pairs.
abstract: 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.
authors.affiliation: University of New Mexico, Albuquerque, NM, USA. twilliams@salud.unm.edu
authors.forename: T M
authors.initials: TM
authors.lastname: Williams
authors.forename: T
authors.initials: T
authors.lastname: Winden
authors.forename: M
authors.initials: M
authors.lastname: Setterholm
authors.forename: C A
authors.initials: CA
authors.lastname: Vierra-Green
authors.forename: S
authors.initials: S
authors.lastname: Spellman
authors.forename: S
authors.initials: S
authors.lastname: Flesch
authors.forename: Z
authors.initials: Z
authors.lastname: Awdeh
authors.forename: L A
authors.initials: LA
authors.lastname: Baxter-Lowe
authors.forename: A B
authors.initials: AB
authors.lastname: Begovich
authors.forename: M
authors.initials: M
authors.lastname: Fernandez-Vina
authors.forename: J
authors.initials: J
authors.lastname: Hegland
authors.forename: C K
authors.initials: CK
authors.lastname: Hurley
authors.forename: D
authors.initials: D
authors.lastname: Johnson
authors.forename: H
authors.initials: H
authors.lastname: Noreen
authors.forename: M
authors.initials: M
authors.lastname: Salazar
authors.forename: B
authors.initials: B
authors.lastname: Schmeckpeper
authors.forename: E J
authors.initials: EJ
authors.lastname: Yunis
mesh_terms: D000483:Alleles; D016026:Bone Marrow Transplantation; D006681:HLA-D Antigens / Q000235:genetics*; D006650:Histocompatibility Testing / Q000379:methods*; D006801:Humans; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D012150:Polymorphism, Restriction Fragment Length; D015203:Reproducibility of Results; D012189:Retrospective Studies; D017422:Sequence Analysis, DNA / Q000379:methods
chemical_list: D006681:HLA-D Antigens; D015345:Oligonucleotide Probes