P027 Automation of NGS-based HLA typing assays on the Beckman Coulter Biomek series of automated liquid handlers

Abstract

The Human Leukocyte Antigen (HLA) system consists of 21 genes and their various alleles (over 12,500 alleles discovered as of January 2015) that allow the immune system to differentiate between native cells and non-native cells and pathogens. Accurate HLA typing is essential for tissue transplants, as a close match between donor and recipient reduces the risk of transplant rejection. Traditionally, HLA typing has been performed with a laborious series of Sanger sequencing based assays that limit sample throughput and sometimes provide ambiguous results due to high levels of homology within the HLA system. More recently, a number of vendors have developed HLA typing assays that utilize next generation sequencing (NGS) technologies. NGS-based HLA typing provides much higher resolution than Sanger sequencing and provides much higher sample throughput due to the massive number of sequencing reads generated in each run. However, preparing DNA samples for HLA typing utilizing NGS-based assays still requires a fairly complicated workflow that involves amplifying the HLA loci and then converting those amplicons to fragment libraries suitable for sequencing. This process necessarily involves a number of pipetting operations that leave open the possibility of pipetting errors, cross-contamination, and errors in maintaining data integrity. To reduce the complexity of the fragment-library preparation, provide greater laboratory efficiency, and prevent errors, we have developed automation protocols from a variety of reagent vendors for NGS-based HLA typing on the Biomek series of automated liquid handlers. These protocols are highly modular in design, allowing for customized installation across one or several automated liquid handlers depending on the laboratory requirements. These automation protocols also utilize Biomek Method Launcher, a new software package that streamlines method operation and instrument setup, as well as HTML-driven user interfaces that allow the user the opportunity to customize their workflows to provide greater efficiency in the laboratory. The Data Acquisition and Reporting Tool (DART) software can also be included to provide improved data handling and reporting to LIMS systems.