A Novel Multiplex droplet digital PCR Assay to Identify and Quantify KRAS Mutations in Clinical Specimens

Publication date: Available online 23 November 2018Source: The Journal of Molecular DiagnosticsAuthor(s): Miguel Alcaide, Matthew Cheung, Kevin Bushell, Sarah E. Arthur, Hui-Li Wong, Joanna Karasinska, Daniel Renouf, David F. Schaeffer, Suzan McNamara, Mathilde Couetoux du Tertre, Gerald Batist, Hagen F. Kennecke, Aly Karsan, Ryan D. MorinRecurrent activating point mutations in KRAS are critical drivers in pancreatic cancer and have been attributed to resistance to anti–epidermal growth factor receptor therapy in colorectal cancer. Although KRAS genotyping provides limited clinical utility in the diagnosis and management of pancreatic cancer patients at present, inferences about the fractional abundance of KRAS mutations may inform on tumor purity in traditionally challenging clinical specimens and their potential use in precision medicine. KRAS genetic testing has indeed become an essential tool to guide treatment decisions in colorectal cancer, but there is an unmet need for methods standardization. Here, we present a unique droplet digital PCR method that enables the simultaneous detection and quantification of KRAS exon 2, 3, and 4 point mutations and copy number alterations. We have validated 13 mutations (G12S, G12R, G12D, G12A, G12V, G12C, G13D, G60V, Q61H, Q61L, A146V, A146T, and A146P) and focal KRAS amplifications by conducting this assay in a cohort of 100 DNA samples extracted from fresh frozen tumor biopsies, formaldehyde-fixed, paraffin-embedded tissue, and li...
Source: The Journal of Molecular Diagnostics - Category: Pathology Source Type: research