title: Automated processing of fluorescence in-situ hybridization slides for HER2 testing in breast and gastro-esophageal carcinomas.
abstract: BACKGROUND
  HER2 fluorescence in-situ hybridization (FISH) is used in breast and gastro-esophageal carcinoma for determining HER2 gene amplification and patients' eligibility for HER2 targeted therapeutics. Traditional manual processing of the FISH slides is labor intensive because of multiple steps that require hands on manipulation of the slides and specifically timed intervals between steps. This highly manual processing also introduces inter-run and inter-operator variability that may affect the quality of the FISH result. Therefore, we sought to incorporate an automated processing instrument into our FISH workflow.
  
  
  METHODS
  Twenty-six cases including breast (20) and gastro-esophageal (6) cancer comprising 23 biopsies and three excision specimens were tested for HER2 FISH (Pathvysion, Abbott) using the Thermobrite Elite (TBE) system (Leica). Up to 12 slides can be run simultaneously. All cases were previously tested by the Pathvysion HER2 FISH assay with manual preparation. Twenty cells were counted by two observers for each case; five cases were tested on three separate runs by different operators to evaluate the precision and inter-operator variability.
  
  
  RESULTS
  There was 100% concordance in the scoring between the manual and TBE methods as well as among the five cases that were tested on three runs. Only one case failed due to poor probe hybridization. In total, seven cases were positive for HER2 amplification (HER2:CEP17 ratio >2.2) and the remaining 19 were negative (HER2:CEP17 ratio <1.8) utilizing the 2007 ASCO/CAP scoring criteria. Due to the automated denaturation and hybridization, for each run, there was a reduction in labor of 3.5h which could then be dedicated to other lab functions.
  
  
  CONCLUSIONS
  The TBE is a walk away pre- and post-hybridization system that automates FISH slide processing, improves work flow and consistency and saves approximately 3.5h of technologist time. The instrument has a small footprint thus occupying minimal counter space. TBE processed slides performed exceptionally well in comparison to the manual technique with no disagreement in HER2 amplification status.
authors.affiliation: Department of Pathology, Dartmouth Hitchcock Medical Center, Lebanon, NH, United States; Norris Cotton Cancer Center, Lebanon, NH, United States; Department of Pathology, Theodor and Audrey Geisel School of Medicine at Dartmouth, Hanover, NH, United States. Electronic address: laura.j.tafe@hitchcock.org.
authors.forename: Laura J
authors.initials: LJ
authors.lastname: Tafe
authors.affiliation: Department of Pathology, Dartmouth Hitchcock Medical Center, Lebanon, NH, United States; Norris Cotton Cancer Center, Lebanon, NH, United States; Department of Pathology, Theodor and Audrey Geisel School of Medicine at Dartmouth, Hanover, NH, United States.
authors.forename: Samantha F
authors.initials: SF
authors.lastname: Allen
authors.affiliation: Department of Pathology, Dartmouth Hitchcock Medical Center, Lebanon, NH, United States; Norris Cotton Cancer Center, Lebanon, NH, United States; Department of Pathology, Theodor and Audrey Geisel School of Medicine at Dartmouth, Hanover, NH, United States.
authors.forename: Heather B
authors.initials: HB
authors.lastname: Steinmetz
authors.affiliation: Department of Pathology, Dartmouth Hitchcock Medical Center, Lebanon, NH, United States; Norris Cotton Cancer Center, Lebanon, NH, United States; Department of Pathology, Theodor and Audrey Geisel School of Medicine at Dartmouth, Hanover, NH, United States.
authors.forename: Betty A
authors.initials: BA
authors.lastname: Dokus
authors.affiliation: Department of Pathology, Dartmouth Hitchcock Medical Center, Lebanon, NH, United States; Norris Cotton Cancer Center, Lebanon, NH, United States; Department of Pathology, Theodor and Audrey Geisel School of Medicine at Dartmouth, Hanover, NH, United States.
authors.forename: Leanne J
authors.initials: LJ
authors.lastname: Cook
authors.affiliation: Department of Pathology, Dartmouth Hitchcock Medical Center, Lebanon, NH, United States; Norris Cotton Cancer Center, Lebanon, NH, United States; Department of Pathology, Theodor and Audrey Geisel School of Medicine at Dartmouth, Hanover, NH, United States.
authors.forename: Jonathan D
authors.initials: JD
authors.lastname: Marotti
authors.affiliation: Department of Pathology, Dartmouth Hitchcock Medical Center, Lebanon, NH, United States; Norris Cotton Cancer Center, Lebanon, NH, United States; Department of Pathology, Theodor and Audrey Geisel School of Medicine at Dartmouth, Hanover, NH, United States.
authors.forename: Gregory J
authors.initials: GJ
authors.lastname: Tsongalis
mesh_terms: D001331:Automation; D001706:Biopsy; D001943:Breast Neoplasms / Q000235:genetics* / Q000473:pathology; D004938:Esophageal Neoplasms / Q000235:genetics* / Q000473:pathology; D005260:Female; D005784:Gene Amplification; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000295:instrumentation* / Q000379:methods*; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000235:genetics*; D015203:Reproducibility of Results; D013274:Stomach Neoplasms / Q000235:genetics* / Q000473:pathology
chemical_list: D018719:Erb-b2 Receptor Tyrosine Kinases; C508053:ERBB2 protein, human
keywords: Automated processing; Breast cancer; Gastric cancer; HER2 FISH; Molecular diagnostic testing; Thermobrite Elite