Design and validation of a next generation sequencing assay for hereditary BRCA1 and BRCA2 mutation testing

PeerJ. 2016 Jun 28:4:e2162. doi: 10.7717/peerj.2162. eCollection 2016.

Abstract

Hereditary breast and ovarian cancer syndrome, caused by a germline pathogenic variant in the BRCA1 or BRCA2 (BRCA1/2) genes, is characterized by an increased risk for breast, ovarian, pancreatic and other cancers. Identification of those who have a BRCA1/2 mutation is important so that they can take advantage of genetic counseling, screening, and potentially life-saving prevention strategies. We describe the design and analytic validation of the Counsyl Inherited Cancer Screen, a next-generation-sequencing-based test to detect pathogenic variation in the BRCA1 and BRCA2 genes. We demonstrate that the test is capable of detecting single-nucleotide variants (SNVs), short insertions and deletions (indels), and copy-number variants (CNVs, also known as large rearrangements) with zero errors over a 114-sample validation set consisting of samples from cell lines and deidentified patient samples, including 36 samples with BRCA1/2pathogenic germline mutations.

Keywords: Analytical validation; BRCA testing; Hereditary breast and ovarian cancer; Next generation sequencing assay.

Grants and funding

The authors received no funding for this work.