Quantitative High-Sensitivity Multiplex Detection of Rare KRAS Mutations through liquid Biopsy using Bio-Rad QX200TM Droplet DigitalTM PCR System
Pancreatic cancer has a survival rate of 9% due to few symptoms being present in its early stages, causing many people who are sick to not be diagnosed until the cancer has gotten sizable or spread outside of the pancreas. To increase the survival rate, early detection of pancreatic cancer is necessary and possible through the use of a liquid biopsy. Circulating tumor DNA (ctDNA) is released from tumor cells and enters the bloodstream, making a liquid biopsy through the use of a blood sample a cheap, non-invasive cancer detection method. However, much of cell-free DNA (cfDNA) is not from a cancer cell and there can be as little as one piece of ctDNA in a single milliliter of plasma. The use of digital droplet PCR (ddPCR) to assess plasma samples has been shown to be effective in monitoring cancer with patients who are metastatic. The experiment conducted optimized the annealing time, annealing temperature, master mix concentration and type via response-surface-methodology (RSM) for a highly sensitive ddPCR assay that will detect KRAS mutations through a liquid biopsy on the Bio-Rad QX200TM Droplet DigitalTM PCR System.