Analytical Evaluation of an NGS Testing Method for Routine Molecular Diagnostics on Melanoma Formalin-Fixed, Paraffin-Embedded Tumor-Derived DNA

摘要: Next Generation Sequencing (NGS) is a promising tool for the improvement of tumor molecular profiling in view identification personalized treatment oncologic patients. To verify potentiality targeted NGS (Ion AmpliSeq™ Cancer Hotspot Panel v2), selected melanoma samples (n = 21) were retrospectively analyzed on S5 platform order to compare performance with conventional techniques adopted our routine clinical setting (Sequenom MassARRAY system, Sanger sequencing, allele-specific real-time PCR). The capability rare and low-frequency mutations main genes involved (BRAF NRAS genes) was verified integrated results deriving from other oncogenes suppressor genes. analytical evaluation carried out by analysis DNA derived control cell lines FFPE (Formalin-Fixed, Paraffin-Embedded) that achieved resolution uncommon variants suitable meet technical requests. Our demonstrate amplicon-based approach can reach sensitivity proper assays together high specificity sequencing method. An overall concordance among tested methods observed classical mutations. assessment quality parameters comparison orthogonal suggest method could be implemented characterization.