The molecular analysis of BRCA1 and BRCA2: Next-generation sequencing supersedes conventional approaches

Valeria D'Argenio, Maria Valeria Esposito, Antonella Telese, Vincenza Precone, Flavio Starnone, Marcella Nunziato, Piergiuseppe Cantiello, Mariangela Iorio, Eloisa Evangelista, Massimiliano D'Aiuto, Alessandra Calabrese, Giulia Frisso, Giuseppe D'Aiuto, Francesco Salvatore

Research output: Contribution to journalArticlepeer-review


Background: Accurate and sensitive detection of BRCA1/2 germ-line mutations is crucial for the clinical management of women affected by breast cancer, for prevention and, notably, also for the identification of at-risk healthy relatives. The most widely used methods for BRCA1/. 2 molecular analysis are Sanger sequencing, and denaturing high performance liquid chromatography (dHPLC) followed by the Sanger method. However, recent findings suggest that next-generation sequencing (NGS)-based approaches may be an efficient tool for diagnostic purposes. In this context, we evaluated the effectiveness of NGS for BRCA gene analysis compared with dHPLC/Sanger sequencing. Methods: Seventy women were screened for BRCA1/2 mutations by both dHPLC/Sanger sequencing and NGS, and the data were analyzed using a bioinformatic pipeline. Results: Sequence data analysis showed that NGS is more sensitive in detecting BRCA1/2 variants than the conventional procedure, namely, dHPLC/Sanger. Conclusion: Next-generation sequencing is more sensitive, faster, easier to use and less expensive than the conventional Sanger method. Consequently, it is a reliable procedure for the routine molecular screening of the BRCA1/2 genes.

Original languageEnglish
Pages (from-to)221-225
Number of pages5
JournalClinica Chimica Acta
Publication statusPublished - 2015


  • BRCA1
  • BRCA2
  • Method comparison in molecular diagnostics
  • Molecular diagnostics
  • Next-generation sequencing

ASJC Scopus subject areas

  • Biochemistry
  • Clinical Biochemistry
  • Biochemistry, medical


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