117-P

Abstract

Aim Currently applied technologies for HLA typing often show ambiguous results. For SBT these are caused by sequencing PCR fragments with a mixture of two alleles. Additional reactions are required to resolve these ambiguities. One of the unique features of Next Generation Sequencing (NGS) is that reads originate from a single molecule. Data obtained by NGS automatically determines separated results of the alleles present. This allows for the development of an HLA genotyping system that yields no ambiguities. In addition, NGS technology is developing fast and has come to a point where it can be applied routinely. We developed NGSengine, a software package to perform HLA typing based on NGS data. In this study we show that HLA typing by NGS can be routinely applied using different NGS systems. Methods NGSengine is an integrated HLA typing approach which includes determining the reference, aligning reads, phasing and typing into one method. Since HLA is highly polymorphic, each of these steps contains HLA-specific features. Data generated from various NGS platforms (Roche/454 GSFLX, Ion Torrent PGM, and Illumina MiSeq) were used to evaluate NGSengine. The sequencing data were kindly provided by D. Monos – Children’s Hospital of Philadelphia. Results All data available from different NGS platforms could be analyzed by NGSengine. In a single analysis the expected HLA typings were obtained. Furthermore results contained no ambiguity or alternative genotypes due to the complete sequencing of the entire HLA gene. Further results will be discussed in detail. Conclusions NGSengine performs reliable HLA typing based on NGS data. NGSengine enables HLA-typing of NGS data in a single procedure. NGSengine can be applied on data from different NGS platforms. Rozemuller: GenDx: Employee; Stockholder. Penning: GenDx: Employee. Mulder: GenDx: Employee; Stockholder.