title: Fully automated fluorescent in situ hybridization (FISH) staining and digital analysis of HER2 in breast cancer: a validation study.
abstract: HER2 assessment is routinely used to select patients with invasive breast cancer that might benefit from HER2-targeted therapy. The aim of this study was to validate a fully automated in situ hybridization (ISH) procedure that combines the automated Leica HER2 fluorescent ISH system for Bond with supervised automated analysis with the Visia imaging D-Sight digital imaging platform. HER2 assessment was performed on 328 formalin-fixed/paraffin-embedded invasive breast cancer tumors on tissue microarrays (TMA) and 100 (50 selected IHC 2+ and 50 random IHC scores) full-sized slides of resections/biopsies obtained for diagnostic purposes previously. For digital analysis slides were pre-screened at 20x and 100x magnification for all fluorescent signals and supervised-automated scoring was performed on at least two pictures (in total at least 20 nuclei were counted) with the D-Sight HER2 FISH analysis module by two observers independently. Results were compared to data obtained previously with the manual Abbott FISH test. The overall agreement with Abbott FISH data among TMA samples and 50 selected IHC 2+ cases was 98.8% (κ = 0.94) and 93.8% (κ = 0.88), respectively. The results of 50 additionally tested unselected IHC cases were concordant with previously obtained IHC and/or FISH data. The combination of the Leica FISH system with the D-Sight digital imaging platform is a feasible method for HER2 assessment in routine clinical practice for patients with invasive breast cancer.
authors.affiliation: Department of Pathology, Pathology Friesland, Leeuwarden, The Netherlands; Department of Pathology and Medical Biology, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands.
authors.forename: Elise M J
authors.initials: EM
authors.lastname: van der Logt
authors.affiliation: Department of Pathology, Pathology Friesland, Leeuwarden, The Netherlands.
authors.forename: Deborah A J
authors.initials: DA
authors.lastname: Kuperus
authors.affiliation: Department of Pathology, Pathology Friesland, Leeuwarden, The Netherlands.
authors.forename: Jan W
authors.initials: JW
authors.lastname: van Setten
authors.affiliation: Department of Pathology, Pathology Friesland, Leeuwarden, The Netherlands.
authors.forename: Marius C
authors.initials: MC
authors.lastname: van den Heuvel
authors.affiliation: Department of Pathology, Isala Klinieken, Zwolle, The Netherlands.
authors.forename: James E
authors.initials: JE
authors.lastname: Boers
authors.affiliation: Department of Pathology and Medical Biology, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands.
authors.forename: Ed
authors.initials: E
authors.lastname: Schuuring
authors.affiliation: Department of Pathology, Pathology Friesland, Leeuwarden, The Netherlands.
authors.forename: Robby E
authors.initials: RE
authors.lastname: Kibbelaar
mesh_terms: D057205:Automation, Laboratory; D001943:Breast Neoplasms / Q000235:genetics* / Q000473:pathology; D005260:Female; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000295:instrumentation* / Q000379:methods; D017404:In Situ Hybridization, Fluorescence / Q000295:instrumentation* / Q000379:methods; D016612:Paraffin Embedding; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000235:genetics*; D015203:Reproducibility of Results; D046888:Tissue Array Analysis
chemical_list: D018719:Erb-b2 Receptor Tyrosine Kinases; C508053:ERBB2 protein, human
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