title: HLA-DRB1 DNA sequencing based typing: an approach suitable for high throughput typing including unrelated bone marrow registry donors.
abstract: The HLA-DRB1 sequencing based typing strategies reported to date require separate amplifications of each sample with a series of group-specific primers followed by sequencing of any resulting polymerase chain reaction (PCR) products. Whilst this results in high resolution typing in the majority of cases, a number of unnecessary amplifications are performed. We report here a novel approach where amplification of the second exon of HLA-DRB1 is performed in a single tube for all alleles. Retrospective analysis of 642 consecutive Western Australian unrelated bone marrow registry donors has shown that this approach results in unambiguous typings in 71.1% of cases. Ambiguities can be readily resolved if necessary with a single additional sequencing reaction on the original PCR product.
authors.affiliation: Department of Clinical Immunology and Biochemical Genetics, Royal Perth Hospital, Perth, Australia. david@dci.uwa.edu.au
authors.forename: D
authors.initials: D
authors.lastname: Sayer
authors.forename: R
authors.initials: R
authors.lastname: Whidborne
authors.forename: B
authors.initials: B
authors.lastname: Brestovac
authors.forename: F
authors.initials: F
authors.lastname: Trimboli
authors.forename: C
authors.initials: C
authors.lastname: Witt
authors.forename: F
authors.initials: F
authors.lastname: Christiansen
mesh_terms: D000483:Alleles; D016026:Bone Marrow Transplantation / Q000276:immunology*; D005190:Family; D015723:Gene Library; D006684:HLA-DR Antigens / Q000235:genetics*; D059811:HLA-DRB1 Chains; D006650:Histocompatibility Testing / Q000379:methods*; D006801:Humans; D016133:Polymerase Chain Reaction; D012042:Registries; D012189:Retrospective Studies; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA / Q000379:methods*; D014019:Tissue Donors
chemical_list: D006684:HLA-DR Antigens; D059811:HLA-DRB1 Chains