TY - JOUR
T1 - Reference standards for gene fusion molecular assays on cytological samples
T2 - an international validation study
AU - Malapelle, Umberto
AU - Pepe, Francesco
AU - Pisapia, Pasquale
AU - Altimari, Annalisa
AU - Bellevicine, Claudio
AU - Brunnström, Hans
AU - Bruno, Rossella
AU - Büttner, Reinhard
AU - Cirnes, Luis
AU - De Andrea, Carlos E.
AU - de Biase, Dario
AU - Dumur, Catherine I.
AU - Lindquist, Kajsa Ericson
AU - Fontanini, Gabriella
AU - Gautiero, Eugenio
AU - Gentien, David
AU - Hofman, Paul
AU - Hofman, Veronique
AU - Iaccarino, Antonino
AU - Lozano, Maria Dolores
AU - Mayo-De-Las-Casas, Clara
AU - Merkelbach-Bruse, Sabine
AU - Pagni, Fabio
AU - Roman, Ruth
AU - Schmitt, Fernando C.
AU - Siemanowski, Janna
AU - Roy-Chowdhuri, Sinchita
AU - Tallini, Giovanni
AU - Tresserra, Francesc
AU - Borght, Sara Vander
AU - Vielh, Philippe
AU - Vigliar, Elena
AU - Vita, Giulia Anna Carmen
AU - Weynand, Birgit
AU - Rosell, Rafael
AU - Vila, Miguel Angel Molina
AU - Troncone, Giancarlo
N1 - Funding Information:
Monitoraggio ambientale, studio ed approfondimento della salute della popolazione residente inaree a rischio—In attuazione della D.G.R. Campanian. 180/2019. POR Campania FESR 2014–2020 Progetto “Sviluppo di Approcci Terapeutici Innovativi per patologie Neoplastiche resistenti ai trattamenti—SATIN”.
Publisher Copyright:
© Author(s) (or their employer(s)) 2023.
PY - 2023/1
Y1 - 2023/1
N2 - Aims Gene fusions assays are key for personalised treatments of advanced human cancers. Their implementation on cytological material requires a preliminary validation that may make use of cell line slides mimicking cytological samples. In this international multi-institutional study, gene fusion reference standards were developed and validated. Methods Cell lines harbouring EML4(13)–ALK(20) and SLC34A2(4)–ROS1(32) gene fusions were adopted to prepare reference standards. Eight laboratories (five adopting amplicon-based and three hybridisation-based platforms) received, at different dilution points two sets of slides (slide A 50.0%, slide B 25.0%, slide C 12.5% and slide D wild type) stained by Papanicolaou (Pap) and May Grunwald Giemsa (MGG). Analysis was carried out on a total of 64 slides. Results Four (50.0%) out of eight laboratories reported results on all slides and dilution points. While 12 (37.5%) out of 32 MGG slides were inadequate, 27 (84.4%) out of 32 Pap slides produced libraries adequate for variant calling. The laboratories using hybridisation-based platforms showed the highest rate of inadequate results (13/24 slides, 54.2%). Conversely, only 10.0% (4/40 slides) of inadequate results were reported by laboratories adopting amplicon-based platforms. Conclusions Reference standards in cytological format yield better results when Pap staining and processed by amplicon-based assays. Further investigation is required to optimise these standards for MGG stained cells and for hybridisation-based approaches.
AB - Aims Gene fusions assays are key for personalised treatments of advanced human cancers. Their implementation on cytological material requires a preliminary validation that may make use of cell line slides mimicking cytological samples. In this international multi-institutional study, gene fusion reference standards were developed and validated. Methods Cell lines harbouring EML4(13)–ALK(20) and SLC34A2(4)–ROS1(32) gene fusions were adopted to prepare reference standards. Eight laboratories (five adopting amplicon-based and three hybridisation-based platforms) received, at different dilution points two sets of slides (slide A 50.0%, slide B 25.0%, slide C 12.5% and slide D wild type) stained by Papanicolaou (Pap) and May Grunwald Giemsa (MGG). Analysis was carried out on a total of 64 slides. Results Four (50.0%) out of eight laboratories reported results on all slides and dilution points. While 12 (37.5%) out of 32 MGG slides were inadequate, 27 (84.4%) out of 32 Pap slides produced libraries adequate for variant calling. The laboratories using hybridisation-based platforms showed the highest rate of inadequate results (13/24 slides, 54.2%). Conversely, only 10.0% (4/40 slides) of inadequate results were reported by laboratories adopting amplicon-based platforms. Conclusions Reference standards in cytological format yield better results when Pap staining and processed by amplicon-based assays. Further investigation is required to optimise these standards for MGG stained cells and for hybridisation-based approaches.
U2 - 10.1136/jclinpath-2021-207825
DO - 10.1136/jclinpath-2021-207825
M3 - Article
C2 - 34429353
AN - SCOPUS:85126172352
SN - 0021-9746
VL - 76
SP - 47
EP - 52
JO - Journal of Clinical Pathology
JF - Journal of Clinical Pathology
IS - 1
ER -
