Digital Sorting of Pure Cell Populations Enables Unambiguous Genetic Analysis of Heterogeneous Formalin-Fixed Paraffin-Embedded Tumors by Next Generation Sequencing

摘要: Precision medicine in oncology requires an accurate characterization of a tumor molecular profile for patient stratification. Though targeted deep sequencing is effective tool to detect the presence somatic sequence variants, significant number specimens do not meet requirements needed routine clinical application. Analysis hindered by contamination normal cells and inherent heterogeneity, compounded with challenges dealing minute amounts tissue DNA damages common formalin-fixed paraffin-embedded (FFPE) specimens. Here we present innovative workflow using DEPArray™ system, microchip-based digital sorter achieve 100%-pure, homogenous subpopulations from FFPE samples. Cells are distinguished fluorescently labeled antibodies content. The ability address heterogeneity enables unambiguous determination true-positive loss-of-heterozygosity as well copy variants. proposed strategy overcomes trade-offs made between sensitivity specificity detecting genetic variants mixed population, thus rescuing analysis even smaller samples low cellularity.