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The Idylla\u2122 GeneFusion Assay detects fusions using fusion-specific as well as expression imbalance detection, the latter enabling detection of uncommon fusions not covered by fusion-specific assays. In total, 326 archival aNSCLC formalin-fixed paraffin-embedded (FFPE) samples were included of which 44% were resected specimen, 46% tissue biopsies, and 9% cytological specimen. With a total of 179 biomarker-positive cases (i.e., 85 <jats:italic>ALK<\/jats:italic>, 33 <jats:italic>ROS1<\/jats:italic>, 20 <jats:italic>RET<\/jats:italic> fusions and 41 <jats:italic>MET<\/jats:italic> exon 14 skipping), this is one of the largest fusion-positive datasets ever tested. The results of the Idylla\u2122 GeneFusion Assay were compared with earlier results of routine reference technologies including fluorescence in situ hybridization, immunohistochemistry, reverse-transcription polymerase chain reaction, and next-generation sequencing, establishing a high sensitivity\/specificity of 96.1%\/99.6% for <jats:italic>ALK<\/jats:italic>, 96.7%\/99.0% for <jats:italic>ROS1<\/jats:italic>, 100%\/99.3% for <jats:italic>RET<\/jats:italic> fusion, and 92.5%\/99.6% for <jats:italic>MET<\/jats:italic> exon 14 skipping, and a low failure rate (0.9%). The Idylla\u2122 GeneFusion Assay was found to be a reliable, sensitive, and specific tool for routine detection of <jats:italic>ALK<\/jats:italic>, <jats:italic>ROS1<\/jats:italic>, <jats:italic>RET<\/jats:italic> fusions and <jats:italic>MET<\/jats:italic> exon 14 skipping. Given its short turnaround time of about 3 h, it is a time-efficient upfront screening tool in FFPE samples, supporting rapid clinical decision making. Moreover, expression-imbalance-based detection of potentially novel fusions may be easily verified with other routine technologies without delaying treatment initiation.<\/jats:p>","DOI":"10.1007\/s00428-024-03778-9","type":"journal-article","created":{"date-parts":[[2024,3,16]],"date-time":"2024-03-16T14:01:35Z","timestamp":1710597695000},"page":"677-686","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Multicenter evaluation of an automated, multiplex, RNA-based molecular assay for detection of ALK, ROS1, RET fusions and MET exon 14 skipping in NSCLC"],"prefix":"10.1007","volume":"484","author":[{"given":"Linea","family":"Melchior","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Astrid","family":"Hirschmann","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paul","family":"Hofman","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christophe","family":"Bontoux","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Angel","family":"Concha","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Salima","family":"Mrabet-Dahbi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pascal","family":"Vannuffel","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emmanuel","family":"Watkin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Martina","family":"Putzov\u00e1","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stefania","family":"Scarpino","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anne","family":"Cayre","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paloma","family":"Martin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Robert","family":"Stoehr","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Arndt","family":"Hartmann","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2024,3,16]]},"reference":[{"issue":"5","key":"3778_CR1","doi-asserted-by":"publisher","first-page":"244","DOI":"10.1002\/gcc.23022","volume":"61","author":"D Kazdal","year":"2022","unstructured":"Kazdal D, Hofman V, Christopoulos P, Ili\u00e9 M, Stenzinger A, Hofman P (2022) Fusion-positive non-small cell lung carcinoma: biological principles, clinical practice, and diagnostic implications. 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The use of these patient samples was approved by the respective local Ethics Committees and was in accordance with the Declaration of Helsinki.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval and consent to participate"}},{"value":"Bontoux Christophe: nothing to declareCayre Anne: nothing to declareConcha Angel: nothing to declareHartmann Arndt: honoraria for lectures or consulting\/advisory boards for Abbvie, Agilent, AstraZeneca, Biocartis, BMS, Boehringer Ingelheim, Cepheid, Diaceutics, Gilead, Illumina, Ipsen, Janssen, Lilly, Merck, MSD, Nanostring, Novartis, Pfizer, Qiagen, QUIP GmbH, Roche, Sanofi, 3DHistotech and other research support from AstraZeneca, Biocartis, Cepheid, Gilead, Illumina, Janssen, Nanostring, Novartis, Owkin, Qiagen, QUIP GmbH, Roche, SanofiHirschmann Astrid: nothing to declareHofman Paul: member of the Biocartis advisory boardMartin Paloma: nothing to declareMelchior Linea: nothing to declareMrabet-Dahbi Salima: nothing to declarePutzov\u00e1 Martina: nothing to declareScarpino Stefania: nothing to declareStoehr Robert: nothing to declareVannuffel Pascal: nothing to declareWatkin Emmanuel: honoraria for lectures or consulting\/advisory boards for AstraZeneca, Janssen, MSD, Incyte, Pierre Fabre","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Conflict of interest"}}]}}