Abstract

Scientific Lecture: SuperSelective Primers for Multiplex Real-time PCR Assays that Assess the Abundance of Rare Mutations Associated with Cancer

Fred Kramer, Professor, New Jersey Medical School Rutgers University

PCR assays are the most rapid, most sensitive, and least expensive way to assess the abundance of mutant DNA fragments present in liquid biopsies.  “SuperSelective” PCR primers, due to their unique design, are extraordinarily specific, able to selectively initiate the synthesis of amplicons on ten mutant DNA fragments in the presence of 1,000,000 wild-type DNA fragments (even though the only difference between the mutant and the wild type is a single-nucleotide polymorphism).  Sets of SuperSelective primers, each possessing unique 5’-tag sequences, enable the amplicons generated from each mutant to be distinguished by differently colored molecular beacon probes.  The inclusion of primers for a wild-type reference gene fragment, enables the abundance of each type of mutant DNA fragment to be assessed by determining the difference between its threshold value and the threshold value of the reference gene.