Combining AmplideX® PCR Chemistries, Nanopore Sequencing, and Automated Analysis Software to Genotype Complex, Clinically-relevant Variants in 11 Challenging Targets Associated with Hereditary Diseases
Summary
• Conventional sequencing methods struggle with
detecting complex variants in key genes
associated with inherited genetic disorders of high
prevalence, necessitating complicated workflows
and producing results that often lack
comprehensive detection of pathogenic variants.
• We developed a novel PCR enrichment/Nanopore
sequencing assay and accompanying analysis
software that detects both simple and complex
variants across CFTR, SMN1/2, FMR1, HBA1/2,
HBB, GBA, CYP21A2, TNXB, and F8 intron
inversions with a streamlined end-to-end
workflow.
• Analysis performance was optimized and
assessed using a mixed sample set consisting of
371 cell-line samples and 141 gDNA samples
isolated from whole blood, which demonstrated
>95% agreement across all variant categories and
genes included in the assay.