Utilizing nanopore sequencing technology for the rapid and comprehensive characterization of eleven HLA loci; addressing the need for deceased donor expedited HLA typing.

Bibliographic Collection: 
CARTA-Inspired Publication
Publication Type: Journal Article
Authors: Mosbruger, Timothy L; Dinou, Amalia; Duke, Jamie L; Ferriola, Deborah; Mehler, Hilary; Pagkrati, Ioanna; Damianos, Georgios; Mbunwe, Eric; Sarmady, Mahdi; Lyratzakis, Ioannis; Tishkoff, Sarah A; Dinh, Anh; Monos, Dimitri S
Year of Publication: 2020
Journal: Hum Immunol
Volume: 81
Issue: 8
Pagination: 413-422
Date Published: 2020 Aug
Publication Language: eng
ISSN: 1879-1166
Keywords: Alleles, Genomics, Genotyping Techniques, High-Throughput Nucleotide Sequencing, Histocompatibility Testing, HLA Antigens, Humans, Nanopore Sequencing, Sequence Analysis, DNA, Tissue Donors
Abstract:

The comprehensive characterization of human leukocyte antigen (HLA) genomic sequences remains a challenging problem. Despite the significant advantages of next-generation sequencing (NGS) in the field of Immunogenetics, there has yet to be a single solution for unambiguous, accurate, simple, cost-effective, and timely genotyping necessary for all clinical applications. This report demonstrates the benefits of nanopore sequencing introduced by Oxford Nanopore Technologies (ONT) for HLA genotyping. Samples (n = 120) previously characterized at high-resolution three-field (HR-3F) for 11 loci were assessed using ONT sequencing paired to a single-plex PCR protocol (Holotype) and to two multiplex protocols OmniType (Omixon) and NGSgo®-MX6-1 (GenDx). The results demonstrate the potential of nanopore sequencing for delivering accurate HR-3F typing with a simple, rapid, and cost-effective protocol. The protocol is applicable to time-sensitive applications, such as deceased donor typings, enabling better assessments of compatibility and epitope analysis. The technology also allows significantly shorter turnaround time for multiple samples at a lower cost. Overall, the nanopore technology appears to offer a significant advancement over current next-generation sequencing platforms as a single solution for all HLA genotyping needs.

DOI: 10.1016/j.humimm.2020.06.004
Alternate Journal: Hum Immunol