Utility of NIST Whole-Genome Reference Materials for the Technical Validation of a Multigene Next-Generation Sequencing Test Bennett O.V. Shum, Ilya.

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Presentation transcript:

Utility of NIST Whole-Genome Reference Materials for the Technical Validation of a Multigene Next-Generation Sequencing Test Bennett O.V. Shum, Ilya Henner, Daniele Belluoccio, Marcus J. Hinchcliffe The Journal of Molecular Diagnostics Volume 19, Issue 4, Pages 602-612 (July 2017) DOI: 10.1016/j.jmoldx.2017.04.004 Copyright © 2017 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Figure 1 Discordant small insertion call at NM_001918.3:c.1210-4delT in reference material 8398. A: In agreement with truth set data, Sanger sequencing reveals a 5T homopolymer in the discordant region (boxed area). B: Visualized in the Integrative Genomics Viewer, validation set data reveal a heterozygous 1-bp T deletion in the discordant region (boxed area). The reference sequence is displayed below the aligned reads. Asterisk indicates a stop codon. The Journal of Molecular Diagnostics 2017 19, 602-612DOI: (10.1016/j.jmoldx.2017.04.004) Copyright © 2017 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Figure 2 Copy number variation detection. Representative relative read depth coverage graphs for 14 exome controls. A: Large duplications were called if relative coverage >1.25 (top dotted line). Sample 15111424 had a duplication within chr2:236949389-236957926. B: Large deletions were called if relative coverage <0.735 (bottom dotted line), and no heterozygous variants were found in the region. Samples 15111203 and 15111508 had a deletion within chr1:206318309-20632548. Data are expressed as means ± SD relative coverage for the genomic region analyzed. The Journal of Molecular Diagnostics 2017 19, 602-612DOI: (10.1016/j.jmoldx.2017.04.004) Copyright © 2017 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Figure 3 Workflow for a methods-based validation for targeted next-generation sequencing (NGS) panels. RM, reference material. The Journal of Molecular Diagnostics 2017 19, 602-612DOI: (10.1016/j.jmoldx.2017.04.004) Copyright © 2017 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions