title: Multicenter Evaluation of a Novel Automated Rapid Detection System of BRAF Status in Formalin-Fixed, Paraffin-Embedded Tissues.
abstract: The mutated BRAF oncogene represents a therapeutic target in malignant melanoma. Because BRAF mutations are also involved in the pathogenesis of other human malignancies, the use of specific BRAF inhibitors might also be extended to other diseases in the future. A prerequisite for the clinical application of BRAF inhibitors is the reliable detection of activating BRAF mutations in routine histopathological samples. In a multicenter approach, we evaluated a novel and fully automated PCR-based system (Idylla) capable of detecting BRAF V600 mutations in formalin-fixed, paraffin-embedded tissue within 90 minutes with high sensitivity. We analyzed a total of 436 samples with the Idylla system. Valid results were obtained in 421 cases (96.56%). Its performance was compared with conventional methods (pyrosequencing or Sanger sequencing). Concordant results were obtained in 406 cases (96.90%). Reanalysis of eight discordant samples by next-generation sequencing and/or pyrosequencing with newly extracted DNA and the BRAF RGQ Kit confirmed the Idylla result in seven cases, resulting in an overall agreement of 98.57%. In conclusion, the Idylla system is a highly reliable and sensitive platform for detection of BRAF V600 mutations in formalin-fixed, paraffin-embedded material, providing an efficient alternative to conventional diagnostic methods, particularly for routine diagnostics laboratories with limited experience in molecular pathology.
authors.affiliation: Department of Pathology, Medical University of Vienna, Vienna, Austria.
authors.forename: Ana-Iris
authors.initials: AI
authors.lastname: Schiefer
authors.affiliation: Institute of Pathology, Charité-Universitaetsmedizin Berlin, Berlin, Germany.
authors.forename: Laura
authors.initials: L
authors.lastname: Parlow
authors.affiliation: Department of Pathology, Medical University of Vienna, Vienna, Austria.
authors.forename: Lisa
authors.initials: L
authors.lastname: Gabler
authors.affiliation: Institute of Pathology, Überlingen, Germany.
authors.forename: Ildiko
authors.initials: I
authors.lastname: Mesteri
authors.affiliation: Department of Pathology, Medical University of Vienna, Vienna, Austria.
authors.forename: Oskar
authors.initials: O
authors.lastname: Koperek
authors.affiliation: Department of Neuropathology, Institute of Pathology, University of Heidelberg, and Clinical Cooperation Unit Neuropathology, DKFZ and DKTK, Heidelberg, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: von Deimling
authors.affiliation: Department of Obstetrics and Gynecology, Medical University of Vienna, Vienna, Austria.
authors.forename: Berthold
authors.initials: B
authors.lastname: Streubel
authors.affiliation: Division of Oncology, Department of Internal Medicine I, Comprehensive Cancer Center Vienna, Medical University of Vienna, Vienna, Austria.
authors.forename: Matthias
authors.initials: M
authors.lastname: Preusser
authors.affiliation: Institute of Pathology, Charité-Universitaetsmedizin Berlin, Berlin, Germany.
authors.forename: Annika
authors.initials: A
authors.lastname: Lehmann
authors.affiliation: Department of Pathology, Johannes Wesling Klinikum Minden, Minden, Germany.
authors.forename: Udo
authors.initials: U
authors.lastname: Kellner
authors.affiliation: Department of Pathology, Antwerp University Hospital, Edegem, Belgium.
authors.forename: Patrick
authors.initials: P
authors.lastname: Pauwels
authors.affiliation: Department of Pathology, Antwerp University Hospital, Edegem, Belgium.
authors.forename: Suzan
authors.initials: S
authors.lastname: Lambin
authors.affiliation: Institute of Pathology, Charité-Universitaetsmedizin Berlin, Berlin, Germany.
authors.forename: Manfred
authors.initials: M
authors.lastname: Dietel
authors.affiliation: Institute of Pathology, Charité-Universitaetsmedizin Berlin, Berlin, Germany. Electronic address: michael.hummel@charite.de.
authors.forename: Michael
authors.initials: M
authors.lastname: Hummel
authors.affiliation: Institute of Pathology, Charité-Universitaetsmedizin Berlin, Berlin, Germany.
authors.forename: Frederick
authors.initials: F
authors.lastname: Klauschen
authors.affiliation: Department of Pathology, Medical University of Vienna, Vienna, Austria. Electronic address: peter.birner@meduniwien.ac.at.
authors.forename: Peter
authors.initials: P
authors.lastname: Birner
authors.affiliation: Institute of Pathology, Charité-Universitaetsmedizin Berlin, Berlin, Germany.
authors.forename: Markus
authors.initials: M
authors.lastname: Möbs
mesh_terms: D004252:DNA Mutational Analysis / Q000379:methods* / Q000592:standards; D005820:Genetic Testing / Q000379:methods* / Q000592:standards; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D008545:Melanoma / Q000175:diagnosis / Q000235:genetics; D025202:Molecular Diagnostic Techniques / Q000379:methods / Q000592:standards; D009154:Mutation; D016612:Paraffin Embedding; D048493:Proto-Oncogene Proteins B-raf / Q000235:genetics*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D016707:Tissue Fixation
chemical_list: D048493:Proto-Oncogene Proteins B-raf