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 se parate amplifications of each sample with a series of group-specific primer s followed by sequencing of any resulting polymerase chain reaction (PCR) p roducts. Whilst this results in high resolution typing in the majority of c ases, a number of unnecessary amplifications are performed Me report here a novel approach where amplification of the second exon of HLA-DRB1 is perfo rmed in a single tube for all alleles. Retrospective analysis of 642 consec utive Western Australian unrelated bone marrow registry donors has shown th at this approach results in unambiguous typings in 71.1% of cases Ambiguiti es can be readily resolved if necessary with a single additional sequencing reaction on the original PCR product.