LAUSR

Aim High-resolution typing results are required to perform stem cell transplantations. This means that for both alleles at least the DNA sequence of core regions exon 2 and 3 for class I loci and exon 2 for class II loci should be determined. However, high resolution typing results are in many cases still ambiguous. Therefore, allelic typing resolution is preferred. Sequencing the complete human leukocyte antigen (HLA) gene always results in one unique genotype. Next-generation sequencing (NGS) enables sequencing whole genes, if needed in high-throughput. Our current NGSgo®-AmpX strategy amplifies the whole gene for class I HLA loci and enables allelic resolution in most cases. For class II HLA loci, high to allelic resolution is obtained depending on the HLA locus analyzed. Here, we will present the results of a novel NGSgo® amplification assay based on full gene amplification of HLA-DRB1. Methods The performance of the NGSgo® Whole Gene assay for HLA-DRB1 has been evaluated on a sample panel of > 90 IHWG and local samples representing different DNA sources. The panel contained all allele groups and their combinations. The amplicons were subsequently tested in the NGSgo® workflow for Illumina (GenDx). Data was analyzed using the HLA typing software package NGSengine® (GenDx). Results Strong amplicons were obtained that showed to be fully compatible with the NGSgo® workflow. The typing results were in concordance with the expected pre-types. Additionally, data analysis showed quality values within the established acceptance criteria. Conclusions The NGSgo®-AmpX Whole Gene amplification strategy for HLA-DRB1 proves to be a reliable, robust and accurate method to obtain allelic typing resolution.