Feasibility of combined detection of gene mutations and fusion transcripts in bone marrow trephines from leukemic neoplasms

Publication date: Available online 7 February 2020Source: The Journal of Molecular DiagnosticsAuthor(s): Stephan Bartels, Britta Hasemeier, Julia Vogtmann, Elisa Schipper, Guntram Büsche, Jerome Schlue, Hans Kreipe, Ulrich LehmannAbstractChromosomal translocations resulting in fusion genes represent important oncogenic drivers and potential therapeutic targets in rare leukemia subtypes. Formalin-fixed and paraffin-embedded trephines are frequently used in hematologic diagnostic and provide relevant access to leukemic cells for further studies, e.g. phenotyping in bone marrow fibrosis. However, high-throughput molecular analysis of nucleic acids obtained from this material is challenging, especially the reliable detection of RNA transcripts.We analyzed 63 formalin-fixed and paraffin-embedded bone marrow trephines of patients with chronic eosinophilic leukemia, chronic myeloid leukemia, acute myeloid leukemia and myeloproliferative neoplasms for gene mutations and the presence of fusion transcripts with a commercial amplicon-based next generation sequencing approach. Thereby we could detect and validate (by reverse transcription PCR) fusion transcripts relevant for diagnosis and therapy in 25 patients (39.7%). Retrospectively selected material, up to 10 years old, was used for this purpose and only one sample failed in the RNA analysis (1.6%).We conclude that amplicon-based fusion transcript detection in bone marrow trephines is feasible and that bone marrow trephines taken fo...
Source: The Journal of Molecular Diagnostics - Category: Pathology Source Type: research