Testing NTRK testing: Wet‐lab and in silico comparison of RNA‐based targeted sequencing assays

• DOI: 10.1002/gcc.22819
• Publication Date: 2019-10-25T16:28:43Z
• Authors: Nicole Pfarr; Martina Kirchner; Ulrich Lehmann; Jonas Leichsenring; Sabine Merkelbach...; Julia Glade; Michael Hummel; Fabian Stögbauer; Annika Lehmann; Marcel Trautmann; Jörg Kumbrink; Andreas Jung; Wolfgang Dietmaier; Volker Endris; Daniel Kazdal; Matthias Evert; David Horst; Hans Kreipe; Thomas Kirchner; Eva Wardelmann; Ulrik Lassen; Reinhard Büttner; Wilko Weichert; Manfred Dietel; Peter Schirmacher; Albrecht Stenzinger
• Keywords: Adult; Male; 0301 basic medicine; Adolescent; Clinical Decision-Making; Receptors, Nerve Growth Factor; FFPE; 03 medical and health sciences; Neoplasms; Biomarkers, Tumor; Humans; Genetic Testing; Child; In Situ Hybridization, Fluorescence; Aged; gene fusion; Disease Management; High-Throughput Nucleotide Sequencing; Infant; Reproducibility of Results; RNA sequencing; Middle Aged; Immunohistochemistry; 3. Good health; NGS; Child, Preschool; Female; NTRK

AbstractNTRK fusions involving three neurotrophic tyrosine receptor kinase genes NTRK1, NTRK2, and NTRK3 and a variety of fusion partners were identified as oncogenic drivers across many cancer types. Drugs that target the chimeric protein product require the identification of the underlying gene fusion. This advocates the diagnostic use of molecular assays ranging from fluorescence in situ hybridization (FISH) and reverse transcription polymerase chain reaction (RT‐PCR)/Sanger approaches to targeted next‐generation sequencing (NGS). Immunohistochemistry may be used as a screening tool and adjunct diagnostic assay in this context. Although FISH and RT‐PCR/Sanger approaches are widely adopted in routine diagnostics, current experience with targeted RNA‐based NGS is limited. Here, we report on the analysis of major assays (TruSight TST170 and TruSight RNA Fusion [Illumina]; Archer FusionPlex Solid Tumor, Archer FusionPlex Lung, and Archer FusionPlex Oncology [Archer]; Oncomine Comprehensive Assay v3 RNA and Oncomine Focus RNA [Thermo Fisher Scientific]) that are commercially available. The data set includes performance results of a multicentric comparative wet‐lab study as well as an in silico analysis on the ability to detect the broad range of NTRK fusions reported until now. A test algorithm that reflects assay methodology is provided. This data will support implementation of targeted RNA sequencing in routine diagnostics and inform screening and testing strategies that have been brought forward.