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 - Ericson Lindquist, Kajsa
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 - Vander Borght, Sara
AU - Vielh, Philippe
AU - Vigliar, Elena
AU - Vita, Giulia Anna Carmen
AU - Weynand, Birgit
AU - Rosell, Rafael
AU - Molina Vila, Miguel Angel
AU - Troncone, Giancarlo
N1 - Publisher Copyright:
© Author(s) (or their employer(s)) 2023. No commercial re-use. See rights and permissions. Published by BMJ.
PY - 2023/1/1
Y1 - 2023/1/1
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.
KW - cytological techniques
KW - molecular
KW - molecular biology
KW - pathology
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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 -