Verification of Nanopore Sequencing Assay for Genetic Variants

This technical report presents the verification of an integrated amplification-based nanopore sequencing assay and automated software solution designed to genotype challenging, clinically relevant genetic variants. The assay addresses the limitations of conventional sequencing methods, which often struggle to detect complex variants associated with inherited genetic disorders. The report outlines the development of a novel PCR enrichment and nanopore sequencing assay that can accurately detect both simple and complex variants across multiple genes. Verification testing was performed using a mixed sample set, demonstrating over 97% agreement across various variant categories. The report details the performance metrics of the assay, including its accuracy in identifying short tandem repeats, copy number variations, and structural variations. Additionally, it describes the streamlined workflow and bespoke software designed to automate analysis, making the characterization of complex genotypes more accessible to laboratories. The findings emphasize the assay's potential for improving diagnostic and screening applications in genetic disorders.