Ultrafast Workflow for Accurate Full-Length ABO Genotyping Using Nanopore Sequencing

Rapid ABO discrepancy resolution is critical for transfusion safety. However, current short-read sequencing cannot resolve phase across the 21.9-kb ABO gene. Here, the aim is to develop a nanopore-based workflow for full-length ABO genotyping. A workflow combining a long-range PCR amplicon spanning the ABO gene was generated from genomic DNA from 63 serologically discrepant donors and sequenced using the Oxford Nanopore Technologies (ONT) Platform. PacBio HiFi sequencing was used as an orthogonal reference standard.