Accessible AmplideX® PCR and Nanopore Sequencing For Rare Variant Characterization in 11 Challenging Genes Associated with Hereditary Diseases
Summary
•Conventional short-read sequencing methods struggle with detecting complex variants in key genes associated with inherited genetic disorders of high prevalence, necessitating multiple complicated workflows which produce results that often lack comprehensive detection of pathogenic variants.
•We developed a prototype assay based on PCR-enrichment, nanopore sequencing, and machine learning models to enable multiplex detection of diverse variant classes including SNVs, INDELs, Exon del/dups, SVs, gene CNVs and STRs in a single workflow.
•We evaluated performance in collaboration with Dr. Lebre at CHU Reims on a set of 155 unique residual clinical samples collected from centers all over France.
•Sample-level genotype agreement was 100% for all samples that passed QC with variants that were either reported or found to be wild-type with no comparator information.