An Amplification-Based, Long-Read Sequencing Assay* Accurately Identifies Clinically-Relevant SNVs, Duplications, and Deletions in Hemoglobinopathy Relevant Genes
Pathogenic thalassemia variants, particularly in HBA1/2, are difficult to detect. Here, we explored whether novel PCR enrichment and nanopore sequencing could detect both large structural variants and SNVs/INDELs in HBA1/2 and HBB simultaneously. Our assay design uses a coverage-based copy number approach to identify large HBA1/2 and HBB variants while also identifying relevant SNVs/INDELs. Large rearrangements and SNVs/INDELs were detected with 100% and >95% sensitivity, respectively, and 100% PPV at the allele level in known pathogenic samples; large HBA1/2 rearrangements were detected with 100% agreement at the sample level during screening.
Authors: Jonathan Turner, Sarah Statt, Ryan Routsong, Chris Fraher, Gary J Latham, and Bradley Hall