Development and validation of a variant detection workflow for BRCA1 and BRCA2 genes and its clinical application based on the Ion Torrent technology

• DOI: 10.1186/s40246-017-0110-x
• Publication Date: 2017-06-26T08:49:07Z
• Authors: Ana Lígia Buzolin; Caroline Mônaco M...; Patricia Rossi Sa...; Andre Yuji Oku; Alexandre Ricardo...; David Santos Marc...; Caio Robledo D An...; Wagner Rosa Baratela; Miguel Mitne-Neto
• Keywords: Ion semiconductor sequencing; Human genetics; Multiplex; Personal genomics

Breast cancer is the most common among women worldwide, and ovarian difficult gynecological tumor to diagnose with lowest chance of cure. Mutations in BRCA1 BRCA2 genes increase risk by 60% breast up 80% women. Molecular tests allow a better orientation for patients carrying these mutations, affecting prophylaxis, treatment, genetic counseling. Here, we evaluated performance panel BRCA2, using Ion Torrent PGM (Life Technologies) platform customized workflow multiplex ligation-dependent probe amplification detection insertions, deletions genes. We validated 26 samples previously analyzed Myriad Genetics Laboratory, our showed 95.6% sensitivity 100% agreement reports, 85% on positive control sample from NIST. also screened 68 clinical found 22 distinct mutations. The selection robust methodology preparation sequencing, together bioinformatics tools optimized data analysis, enabled development very sensitive test high reproducibility. highlight need explore limitations NGS technique strategies overcome them clinically confident manner.