HLA-DRB1 DNA sequencing based typing: an approach suitable for high throughput typing including unrelated bone marrow registry donors.
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.
Sayer
D
D
Department of Clinical Immunology and Biochemical Genetics, Royal Perth Hospital, Perth, Australia. david@dci.uwa.edu.au
Whidborne
R
R
Brestovac
B
B
Trimboli
F
F
Witt
C
C
Christiansen
F
F
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
D006684:HLA-DR Antigens; D059811:HLA-DRB1 Chains