Lab Med Online.  2021 Oct;11(4):283-289. 10.47429/lmo.2021.11.4.283.

Evaluation of a Targeted Next-generation Sequencing Assay for BRCA Mutation Screening in Clinical Samples

• Kim MJ
• Song BJ
• Kim SE
• Lee CS
• Im JE
• Oh E
• Lee K
• Cho SI
• Kim KJ
• Park SS
• Seong MW

• KMID: 2526090
• DOI: http://doi.org/10.47429/lmo.2021.11.4.283

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 BRCAaccuTest^TM (NGeneBio, Korea).
Methods
BRCAaccuTest^TM 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 BRCAaccuTest^TM in identifying BRCA1/2mutations 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 BRCAaccuTest^TM to detect the BRCA1/2 mutations with high accuracy.

Keyword

BRCA; Mutation; Next-generation sequencing; in vitro diagnosis; Hereditary breast and ovarian cancer (HBOC) syndrome