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Targeted exome sequencing designed for blood group, platelet, and neutrophil antigen investigations: Proof-of-principle study for a customized single-test system


Background Immunohematology reference laboratories provide red blood cell (RBC), platelet (PLT), and neutrophil typing to resolve complex cases, using serology and commercial DNA tests that define clinically important antigens. Broad-range exome sequencing panels that include blood group targets provide accurate blood group antigen predictions beyond those defined by serology and commercial typing systems and identify rare and novel variants. The aim of this study was to design and assess a panel for targeted exome sequencing of RBC, PLT, and neutrophil antigen–associated genes to provide a comprehensive profile in a single test, excluding unrelated gene targets. Study Design and Methods An overlapping probe panel was designed for the coding regions of 64 genes and loci involved in gene expression. Sequencing was performed on 34 RBC and 17 PLT/neutrophil reference samples. Variant call outputs were analyzed using software to predict star allele diplotypes. Results were compared with serology and previous sequence genotyping data. Results Average coverage exceeded 250×, with more than 94% of targets at Q30 quality or greater. Increased coverage revealed a variant in the Scianna system that was previously undetected. The software correctly predicted allele diplotypes for 99.5% of RBC blood groups tested and 100% of PLT and HNA antigens excepting HNA-2. Optimal throughput was 12 to 14 samples per run. Conclusion This single-test system demonstrates high coverage and quality, allowing for the detection of previously overlooked variants and increased sample throughput. This system has the potential to integrate genomic testing across laboratories within hematologic reference settings.

Type Journal
ISBN 0041-1132
Authors Roulis, Eileen; Schoeman, Elizna; Hobbs, Matthew; Jones, Greg; Burton, Mark; Pahn, Gail; Liew, Yew-Wah; Flower, Robert; Hyland, Catherine
Responsible Garvan Author Dr Matthew Hobbs
Publisher Name TRANSFUSION
Published Date 2020-09-30
Published Volume 60
Published Issue 9
Published Pages 2108-2120
Status Published in-print
DOI 10.1111/trf.15945
URL link to publisher's version