Evaluation of a Targeted Next-generation Sequencing Assay for BRCA Mutation Screening in Clinical Samples
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Background: While several factors contribute to breast cancer pathogenesis, hereditary breast cancer results from a genetic predisposition. Genes associated with hereditary breast cancer may be divided into high- and low-penetrance genes depending on their risk rates. BRCA1 and BRCA2 are typical high-penetrance genes that increase the risk of developing breast and ovarian cancers upon undergoing mutations. This study aimed to evaluate the clinical performance of BRCAaccuTestTM (NGeneBio, Korea). Methods: BRCAaccuTestTM is a reagent used to produce libraries for analyzing BRCA1/2 genes using next-generation sequencing (NGS), which analyzes blood-derived genomic DNA. Libraries with adapters and barcodes compatible with the Illumina platform were produced. The clinical performance of NGS-based BRCAaccuTestTM in identifying BRCA1/2 mutations was compared with that of the traditional Sanger sequencing method. Both NGS and Sanger sequencing were performed in a single laboratory using archival DNA from blood samples of 212 patients with breast cancer. Results: All target regions amplified were successfully sequenced to obtain a minimum coverage of 20, demonstrating 100% concordance with the pathogenic single-nucleotide variations and small insertions-deletions previously identified by Sanger sequencing. Conclusions: This study demonstrates the feasibility of using BRCAaccuTestTM to detect the BRCA1/2 mutations with high accuracy.
Abstract
Background: While several factors contribute to breast cancer pathogenesis, hereditary breast cancer results from a genetic predisposition. Genes associated with hereditary breast cancer may be divided into high- and low-penetrance genes depending on their risk rates. BRCA1 and BRCA2 are typical high-penetrance genes that increase the risk of developing breast and ovarian cancers upon undergoing mutations. This study aimed to evaluate the clinical performance of BRCAaccuTestTM (NGeneBio, Korea). Methods: BRCAaccuTestTM is a reagent used to produce libraries for analyzing BRCA1/2 genes using next-generation sequencing (NGS), which analyzes blood-derived genomic DNA. Libraries with adapters and barcodes compatible with the Illumina platform were produced. The clinical performance of NGS-based BRCAaccuTestTM in identifying BRCA1/2 mutations was compared with that of the traditional Sanger sequencing method. Both NGS and Sanger sequencing were performed in a single laboratory using archival DNA from blood samples of 212 patients with breast cancer. Results: All target regions amplified were successfully sequenced to obtain a minimum coverage of 20, demonstrating 100% concordance with the pathogenic single-nucleotide variations and small insertions-deletions previously identified by Sanger sequencing. Conclusions: This study demonstrates the feasibility of using BRCAaccuTestTM to detect the BRCA1/2 mutations with high accuracy.
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