title: Resolution of HLA class I sequence-based typing ambiguities by group-specific sequencing primers.
abstract: The increasing demand for allele-level human leukocyte antigen (HLA) typing has led the sequence-based typing (SBT) to become the preferred method. In turn, the steady increase in the number of HLA alleles driven by the adoption of SBT as the ultimate typing method leads to the ever increasing number of cis/trans ambiguities. Over the last few years, additional sequencing with the commercially available group-specific sequencing primers (GSSPs) has replaced sequence-specific primer-polymerase chain reaction and group-specific amplification as the means of resolving cis/trans ambiguities in many laboratories. Here we summarize our 3-year experience in designing and utilizing GSSPs for resolution of HLA class I ambiguities. The panel of GSSPs used in our laboratory includes 14 primers for HLA-A, 18 for HLA-B, and 13 primers for HLA-C. The panel resolves 99.9% of all ambiguities.
authors.affiliation: HLA Laboratory, American Red Cross Blood Services, New England Region, Dedham, MA 02026, USA. LebedevaT@usa.redcross.org
authors.forename: T V
authors.initials: TV
authors.lastname: Lebedeva
authors.forename: S A
authors.initials: SA
authors.lastname: Mastromarino
authors.forename: E
authors.initials: E
authors.lastname: Lee
authors.forename: M
authors.initials: M
authors.lastname: Ohashi
authors.forename: S M
authors.initials: SM
authors.lastname: Alosco
authors.forename: N
authors.initials: N
authors.lastname: Yu
mesh_terms: D000483:Alleles; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D015343:DNA Probes, HLA / Q000032:analysis / Q000235:genetics; D005189:False Positive Reactions; D005805:Genes, MHC Class I / Q000235:genetics*; D006650:Histocompatibility Testing / Q000379:methods*; D006801:Humans; D008969:Molecular Sequence Data; D012189:Retrospective Studies; D017422:Sequence Analysis, DNA / Q000379:methods / Q000592:standards*; D017385:Sequence Homology; D012984:Software; D013379:Substrate Specificity / Q000235:genetics / Q000276:immunology
chemical_list: D017931:DNA Primers; D015343:DNA Probes, HLA