A Novel Multiplex Droplet Digital PCR Assay to Identify and Quantify KRAS Mutations in Clinical Specimens
Autor: | Ryan D. Morin, Sarah E. Arthur, Daniel J. Renouf, Hui-Li Wong, David F. Schaeffer, Miguel Alcaide, Suzan McNamara, Kevin Bushell, Matthew C. Cheung, Mathilde Couetoux du Tertre, Gerald Batist, Joanna M. Karasinska, Hagen F. Kennecke, Aly Karsan |
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Rok vydání: | 2018 |
Předmět: |
0301 basic medicine
Genotype Colorectal cancer medicine.disease_cause Pathology and Forensic Medicine Proto-Oncogene Proteins p21(ras) 03 medical and health sciences 0302 clinical medicine Pancreatic cancer Medicine Humans Digital polymerase chain reaction Multiplex Liquid biopsy Genotyping business.industry Point mutation DNA Neoplasm Exons medicine.disease 3. Good health Pancreatic Neoplasms 030104 developmental biology 030220 oncology & carcinogenesis Mutation Cancer research Molecular Medicine KRAS business Colorectal Neoplasms Multiplex Polymerase Chain Reaction |
Zdroj: | The Journal of molecular diagnostics : JMD. 21(2) |
ISSN: | 1943-7811 |
Popis: | Recurrent 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 an unmet need for methods standardization exists. 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 liquid biopsy specimens. Despite its modest lower limit of detection (approximately 1%), this assay will be a rapid cost-effective means to infer the purity of biopsy specimens carrying KRAS mutations and can be used in noninvasive serial monitoring of circulating tumor DNA to evaluate clinical response and/or detect early signs of relapse. |
Databáze: | OpenAIRE |
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