An optimized workflow to evaluate estrogen receptor gene mutations in small amounts of cell-free DNA

Publication date: Available online 6 October 2018Source: The Journal of Molecular DiagnosticsAuthor(s): Silvia R. Vitale, Anieta M. Sieuwerts, Nick Beije, Jaco Kraan, Lindsay Angus, Bianca Mostert, Esther A. Reijm, Ngoc M. Van, Ronald van Marion, Luc Y. Dirix, Paul Hamberg, Felix E. de Jongh, Agnes Jager, John A. Foekens, Paolo Vigneri, Stefan Sleijfer, Maurice P.H.M. Jansen, John W.M. MartensThe detection of mutated genes in cell-free DNA (cfDNA) in plasma has emerged as an important minimally-invasive way to obtain detailed information regarding tumor biology. Reliable determination of circulating tumor–derived DNA, often present at a very low quantity amidst an excess of normal DNA in plasma, would be of added value for screening and monitoring of cancer patients and for hypothesis-generating studies in valuable retrospective cohorts. Our aim was to establish a workflow to simultaneously assess four hotspot estrogen receptor mutations (mESR1) in cfDNA isolated from only 200 μL of plasma by means of uniplex or multiplex pre-amplification combined with digital PCR (dPCR). This workflow was then applied in metastatic breast cancer (MBC) patients receiving systemic therapies for MBC. In accordance with previous studies, mESR1 were more frequently detected in endocrine-treated MBC patients at progressive disease (34.1% (15/44)) than before the start of endocrine therapy (3.9% (2/51); P = 0.001). For a subset of samples, results were compared with analysis of these mutations ...
Source: The Journal of Molecular Diagnostics - Category: Pathology Source Type: research