Alexandra S Whale
Molecular and Cell Biology Team, UK
Title: Detection of rare sequence variants using digital PCR
Biography
Biography: Alexandra S Whale
Abstract
Rare mutation detection is offering increasingly important potential in cancer management, the detection of antimicroibal resistance and in monitoring organ transplant rejection. The main challenge is detecting the rare mutant due to background signal from cross reactvity with the predominant wild type sequences. Digital PCR (dPCR) is the latest incarnation of PCR that subdivides the reaction across a large number of smaller reactions (termed ‘partitions’) so that some contains no template molecules. PCR is then performed with fluorescent probes to generate partitions with either an amplification signal or no amplification signal, hence the name ‘digital’. While this subdivision increases the precision of dPCR over qPCR, it also improves the detection of rare mutant variants, which in a normal qPCR would be lost within the high wild type background. Using a range of different models, we are investigating the ability of dPCR to detect rare mutations in DNA and RNA templates. This presentation will demonstrate the ability of dPCR to detect a mutation in less than 1% of the population and to highlight the many factors that can confound this method such as reduced template concentration and competitive PCR.