Automation of ALK gene rearrangement testing with fluorescence in situ hybridization (FISH): A feasibility study

Karen Zwaenepoel (1) , Dennis Merkle (2) , Florian Cabillic (3, 4) , Erica Berg (5) , Marc-Antoine Belaud-Rotureau (4, 3) , Vittorio Grazioli (6) , Olga Herelle (7) , Michael Hummel (5) , Michele Le Calve (4) , Dido Lenze (5) , Stefanie Mende (5) , Patrick Pauwels (1) , Benoit Quilichini (8) , Elena Repetti (6)

1 Department of Pathology
2 Abbott Molecular, Germany
3 Foie, métabolismes et cancer
4 Service de Cytogénétique et de Biologie Cellulaire
5 Charité - UniversitätsMedizin = Charité - University Hospital [Berlin]
6 Laboratoire de biologie moléculaire et génétique
7 Abbott Molecular, Paris
8 Biomnis

In the past several years we have observed a significant increase in our understanding of molecular mechanisms that drive lung cancer. Specifically in the non-small cell lung cancer sub-types, ALK gene rearrangements represent a sub-group of tumors that are targetable by the tyrosine kinase inhibitor Crizotinib, resulting in significant reductions in tumor burden. Phase II and III clinical trials were performed using an ALK break-apart FISH probe kit, making FISH the gold standard for identifying ALK rearrangements in patients. FISH is often considered a labor and cost intensive molecular technique, and in this study we aimed to demonstrate feasibility for automation of ALK FISH testing, to improve laboratory workflow and ease of testing. This involved automation of the pre-treatment steps of the ALK assay using various protocols on the VP 2000 instrument, and facilitating automated scanning of the fluorescent FISH specimens for simplified enumeration on various backend scanning and analysis systems. The results indicated that ALK FISH can be automated. Significantly, both the Ikoniscope and BioView system of automated FISH scanning and analysis systems provided a robust analysis algorithm to define ALK rearrangements. In addition, the BioView system facilitated consultation of difficult cases via the internet.

Mots clés: Fluorescence in situ hybridization; FISH; VP 2000; Automation; BioView; ALK

Domaines: Sciences du Vivant [q-bio]; Cancer

Karen Zwaenepoel, Dennis Merkle, Florian Cabillic, Erica Berg, Marc-Antoine Belaud-Rotureau, et al.. Automation of ALK gene rearrangement testing with fluorescence in situ hybridization (FISH): A feasibility study. Experimental and Molecular Pathology, 2015, 98 (1), pp.113-118. ⟨10.1016/j.yexmp.2015.01.005⟩. ⟨hal-01116397⟩