KRAS Mutation Detection in Oligometastatic Tumors

KRAS mutations were assessed on formalin-fixed paraffin-embedded (FFPE) primary and metastatic tumor tissues from oligometastatic patients. The DNA was extracted from three 10-µm FFPE sections through the MGF03-Genomic DNA FFPE One-Step Kit, according to the manufacturer’s protocol (MagCore Diatech). DNA quality was established in triplicate using the FFPE QC Kit according to the manufacturer’s protocol (Illumina, San Diego, USA). The libraries were prepared with TruSigt TMOncology 500 kit. Sequencing was performed on an Illumina NovaSeq 6000 (San Diego, USA) platform. The assay detects small nucleotide variants (SNVs), indels, splice variants, and immunotherapy biomarkers in 523 cancer-relevant genes. However, our analysis focused on KRAS-related genetic results. The Illumina TruSighth Oncology 500 bioinformatics pipeline was applied to analyze quantitatively and qualitatively the sequencing results, as previously reported (3 (link), 4 (link)).