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Exp Mol Pathol. 2015 Feb;98(1):113-8. doi: 10.1016/j.yexmp.2015.01.005. Epub 2015 Jan 8.

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

Author information

1
Antwerp University Hospital, Department of Pathology, Wilrijkstraat 10, 2650 Edegem, Belgium. Electronic address: Karen.Zwaenepoel@uza.be.
2
Abbott Molecular, Max-Planck-Ring 2, 65205 Wiesbaden, Germany.
3
University of Rennes 1, Faculty of Medicine, Service de Cytogénétique et Biologie Cellulaire, CHU de Rennes, rue Henri Le Guilloux, 35033 Rennes CEDEX 9, France.
4
Charite University Hospital Berlin, Campus Benjamin Franklin, Hindenburgdamm 30, 12200 Berlin, Germany.
5
Italian Center of Diagnostics, Laboratory of Molecular Biology and Genetics, Via Saint Bon 20, 20147 Milan, Italy.
6
Abbott Molecular, Rue de la Couture 12, 94518 Rungis, France.
7
Antwerp University Hospital, Department of Pathology, Wilrijkstraat 10, 2650 Edegem, Belgium.
8
Biomnis Laboratory, Laboratory of Cytogenetics, 17-19 av Tony Garnier, 69357 Lyon, France.

Abstract

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.

KEYWORDS:

ALK; Automation; BioView; FISH; Fluorescence in situ hybridization; VP 2000

PMID:
25576649
DOI:
10.1016/j.yexmp.2015.01.005
[Indexed for MEDLINE]
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