BRCAssure®: BRCA1 and BRCA2 Comprehensive Analysis

Next-generation sequencing: Genomic regions of interest are selected using a custom capture reagent for target enrichment and sequenced via the Illumina® next generation sequencing platform. Regions of interest include all exons and intron/exon junctions (+/-20 nucleotides) of the BRCA1 (NM_007294.3) and BRCA2 (NM_000059.3) genes. Certain regions may be extended to include non-coding sequences known to harbor pathogenic/likely pathogenic variants. Sequencing reads are aligned with the human genome reference GRCh37/hg19 build. Minimum mean coverage is 40X. Any segment failing minimum read depth coverage is rescued by bi- directional Sanger sequencing to complete sequence analysis. Variants, including SNVs and CNVs, are identified using a custom bioinformatics pipeline.

Reported variants: Pathogenic and likely pathogenic variants and variants of uncertain significance (VUS) are reported. Non-deletion variants are specified using the numbering and nomenclature recommended by the Human Genome Variation Society (HGVS). Benign variants are not reported. Variant classification and confirmation are consistent with ACMG standards and guidelines. ^2,3 Detailed variant classification information is available upon request. A variant of uncertain significance (VUS) should not be used in clinical decision making; a VUS is classified based on inadequate or conflicting evidence regarding its pathogenicity or clinical relevance.