Characterisation and Confirmation of new HLA-Alleles found by DKMS Life Science Lab. Novel HLA-Alleles

The high-throughput department of DKMS Life Science Lab encounters novel HLA alleles on a daily basis. To characterise these alleles, we have developed a system to sequence the whole gene from 5’-UTR to 3’-UTR for the HLA loci A, B, C, DQB1 and DPB1 for submission to the European Molecular Biology Laboratory - European Nucleotide Archive (EMBL-ENA) and the IPD-IMGT/HLA Database. Our workflow is based on a dual redundant sequencing strategy. Using shotgun sequencing on an Illumina MiSeq instrument and Single Molecule Real-Time (SMRT) sequencing on a PacBio® RS II instrument, we are able to achieve highly accurate HLA full-length consensus sequences. Remaining conflicts are resolved using the R package DR2S (Dual Redundant Reference Sequencing). Given the relatively high throughput of this strategy, we have developed the semi-automated web service TypeLoader, to aid in the submission of sequences to the EMBL-ENA and the IPD-IMGT/HLA Database.