High Sensitivity and Specificity NGS Analysis of Prostate and Kidney Cancer Samples Using a Novel Liquid Biopsy Workflow.

e16607Background: Next generation sequencing (NGS) of blood-derived nucleic acids is an emerging paradigm for determining the mutational status of cancer patients over time. Circulating tumor cells (CTC) and cell-free circulating DNA have been proposed as possible sample types. Here we present data from a CTC enrichment modality that results in tumor cell purities of > 10% and a high sensitivity NGS data analysis workflow that enables the use of standard amplicon panels typically used for primary tissue. This study was aimed at urological cancers of the kidney and prostate). Methods: Blood samples from prostate and kidney cancer patients were enriched for CTCs using the IsoFlux System, from a starting blood volume of 7.5-14ml. Matched samples were enumerated to determine the CTC load, where CTCs were defined as CK+, CD45- nucleated cells (DAPI+). Cells were lysed and DNA was amplified by whole genome amplification (WGA) using the NGS Kit (Fluxion), and quantified via qPCR. Targeted libraries were sequence...