Evaluation of a Next-Generation Sequencing Assay for BRCA1 and BRCA2 Mutation Detection
Gabriele Lorenzo Capone1, Anna Laura Putignano1, Sharon Trujillo Saavedra1
1Department of Experimental and Clinical Biomedical Sciences Mario Serio, Medical Genetics Unit, University of Florence, Florence, Italy.
The Journal of Molecular Diagnostics : JMD
|October 25, 2017
Summary
The Devyser BRCA next-generation sequencing (NGS) kit accurately detects genetic variants in BRCA1 and BRCA2 genes. This validated assay demonstrates high concordance, making it suitable for clinical diagnostics of hereditary breast and ovarian cancer.
Area of Science:
- Molecular Biology
- Genetics
- Oncology
Background:
- Accurate detection of BRCA1 and BRCA2 gene mutations is crucial for diagnosing and managing hereditary breast and ovarian cancer risk.
- Next-generation sequencing (NGS) offers a comprehensive approach for analyzing these high-risk genes.
Purpose of the Study:
- To evaluate the diagnostic efficiency of the Devyser BRCA kit, a novel targeted NGS assay.
- To assess the kit's capability in detecting various genetic alterations, including nucleotide substitutions, small and large deletions/duplications in BRCA1 and BRCA2 genes.
- To determine the false-negative and false-positive rates of the assay in a clinical laboratory setting.
Main Methods:
- Targeted NGS using the Devyser BRCA kit was performed on 48 samples with known variants (444 small variants, 7 gross rearrangements).
- Assay performance was validated by comparing NGS results against gold standards.
- An additional 179 samples were prospectively analyzed, with variants detected by NGS confirmed using Sanger sequencing or multiplex ligation probe amplification.
Main Results:
- The Devyser BRCA kit demonstrated 100% concordance with gold standards for previously identified variants.
- All 43 additionally detected variants of potential clinical significance in the prospective set were confirmed by orthogonal methods.
- The assay successfully identified nucleotide substitutions, small deletions/insertions, and large deletions/duplications across all 48 coding exons of BRCA1 and BRCA2.
Conclusions:
- The Devyser BRCA kit is an efficient and accurate tool for comprehensive analysis of BRCA1 and BRCA2 genes.
- The assay exhibits satisfactory performance, including low false-negative and false-positive rates, for clinical laboratory use.
- This NGS test provides reliable detection of diverse genetic alterations relevant to hereditary breast and ovarian cancer.
