Assessing the necessity of confirmatory testing for exome-sequencing results in a clinical molecular diagnostic laboratory

Purpose: Sanger sequencing is currently considered the gold standard methodology for clinical molecular diagnostic testing. However, next-generation sequencing has already emerged as a much more efficient means to identify genetic variants within gene panels, the exome, or the genome. We sought to assess the accuracy of next! generation sequencing variant identification in our clinical genomics t laboratory with the goal of establishing a quality score threshold for confirmatory-Sanger-based testing. Methods: Confirmation data for reported results from 144 sequential clinical exome-sequencing cases (94 unique variants) and an additional set of 16 variants from comparable research samples were analyzed. Results: Of the 110 total single-nucleotide variants analyzed, 103 variants had a quality score Q500,103 (100%) of which were confirmed by Sanger sequencing. Of the remaining seven variants with quality scores