Clinical validation of the next-generation sequencing-based Extended RAS Panel assay using metastatic colorectal cancer patient samples from the phase 3 PRIME study

摘要: To validate a next-generation sequencing (NGS)-based companion diagnostic using the MiSeqDx® instrument to simultaneously detect 56 RAS mutations in DNA extracted from formalin-fixed paraffin-embedded metastatic colorectal cancer (mCRC) tumor samples PRIME study. The test’s ability identify patients with mCRC likely benefit panitumumab treatment was assessed. Samples PRIME, which compared first-line panitumumab + FOLFOX4 FOLFOX4, were processed according predefined criteria multiplex assay that included input qualification, library preparation, sequencing, and bioinformatics reporting pipeline. NGS mutational analysis of KRAS NRAS exons 2, 3, 4 performed Sanger sequencing. In 441 samples, positive percent agreement Extended Panel 98.7% negative 97.6%. For clinical validation (n = 528), progression-free survival (PFS) overall (OS) between (RAS Positive) those without Negative). Panitumumab + FOLFOX4 improved PFS Negative (P = 0.02). Quantitative interaction testing indicated effect (measured by hazard ratio versus FOLFOX4) differed for Positive (P = 0.0038) OS (P = 0.0323). allows broad, rapid, highly specific analyses genomic regions. These results support use as selecting panitumumab, utilization is consistent recent guidelines regarding testing. Overall, approximately 13% more detected exon 2 alone. NCT00364013 (ClinicalTrials.gov).