Rapid, scalable and highly automated HLA genotyping using next-generation sequencing: a transition from research to diagnostics

摘要: Human leukocyte antigen matching at allelic resolution is proven clinically significant in hematopoietic stem cell transplantation, lowering the risk of graft-versus-host disease and mortality. However, due to ever growing HLA allele database, tissue typing laboratories face substantial challenges. In light complexity high degree diversity, it has become increasingly difficult define classical transplantation antigens high-resolution by using well-tried methods. Thus, next-generation sequencing entering into diagnostic perfect time serving as a promising tool overcome intrinsic problems. Therefore, we have developed validated scalable automated class I II approach suitable for use. A validation panel 173 clinical proficiency testing samples was analysed, demonstrating 100% concordance reference method. From total 1,273 loci were able generate 1,241 (97.3%) initial successful typings. The mean ambiguity reduction analysed 93.5%. Allele assignment including intronic sequences showed an improved (99.2%) non-expressed alleles. We provide powerful protocol offering short turnaround only two days, fully integrated workflow most importantly reliability. presented assay flexible can be scaled specific primer compilations use different 454 systems. successfully according policies European Federation Immunogenetics. Next-generation seems one new methods field Histocompatibility.