153-P: HIGH-RESOLUTION TYPING BY NEXT-GENERATION SEQUENCING IN A CLINICAL HLA LABORATORY

Abstract

The aim of this study is to test the feasibility and compare the analytical performance of HLA typing of clinical and proficiency testing samples using next-generation sequencing against standard molecular HLA typing methods in a medium-sized clinical HLA laboratory without prior sequencing experience. We amplified HLA-A, -B, -C, -DRB1 and –DQB1 genes, from clinical samples, by long-range PCR using primers developed at our laboratory. Long-range PCR product identity was verified by Luminex SSO. HLA gene products were pooled and processed to prepare a sequencing library using the Nextera XT sample kit followed by sequencing on the Miseq instrument. Data was analyzed using the Omixon Target HLA software to assign HLA types. HLA assignment by next-generation sequencing was compared against results previously obtained by standard molecular typing methods. We report the concordance of HLA typing results obtained by next-generation sequencing and analysis compared with standard molecular typing methodologies. Importantly, the reagent cost of performing high-resolution typing by next-generation sequencing is similar to molecular methods currently used. Next generation sequencing can be a viable approach to high resolution HLA typing in medium-sized clinical HLA laboratories. Berces: Omixon: Employee; Stockholder. Major: Omixon: Employee. Hague: Omixon: Employee.