HLA-DRB1 DNA sequencing based typing: an approach suitable for high throughput typing including unrelated bone marrow registry donors

Authors


Correspondence to:David Sayer
Department of Clinical Immunology and Biochemical Genetics
Royal Perth Hospital
GPO Box X2213
Perth 6001
Australia
Tel: +61 8 92242899
Fax: +61 8 92242920
e-mail: david@dci.uwa.edu.au

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.

Ancillary