Abstract

Breakfast Lecture: Multiplex Real-Time PCR Assays that Assess the Abundance of Rare Somatic Mutations Associated with Cancer

Fred Kramer, Professor of Microbiology, Biochemistry & Molecular Genetics, New Jersey Medical School Rutgers University

“SuperSelective” PCR primers, due to their unique design, exponentially amplify rare mutant DNA fragments present in a sample, while ignoring abundant related wild-type DNA fragments, even if the only difference between the mutant and the wild-type is a single-nucleotide polymorphism.  Different target mutations whose presence suggests the same therapeutic action can be detected with molecular beacons bearing the same colored fluorophore.  Differently colored molecular beacons are simultaneously present in these assays to determine which therapeutic action should be taken and which therapeutic action should be discontinued.  These multiplex, homogeneous assays are sensitive, rapid, inexpensive, utilize existing PCR instruments, and can be carried out frequently during the course of treatment utilizing non-invasive liquid biopsy samples.