title: Standardization of HER2 immunohistochemistry in breast cancer by automated quantitative analysis.
abstract: BACKGROUND
  There is critical need for standardization of HER2 immunohistochemistry testing in the clinical laboratory setting. Recently, the American Society of Clinical Oncology and the College of American Pathologists have submitted guidelines recommending that laboratories achieve 95% concordance between assays and observers for HER2 testing.
  
  
  OBJECTIVE
  As a potential aid to pathologists for achieving these new guidelines, we have conducted an examination using automated quantitative analysis (AQUA analysis) to provide a standardized HER2 immunohistochemistry expression score across instruments (sites), operators, and staining runs.
  
  
  METHODS
  We analyzed HER2 expression by immunohistochemistry in a cohort (n = 669) of invasive breast cancers in tissue microarray format across different instruments (n = 3), operators (n = 3), and staining runs (n = 3). Using light source, instrument calibration techniques, and a new generation of image analysis software, we produced normalized AQUA scores for each parameter and examined their reproducibility.
  
  
  RESULTS
  The average percent coefficients of variation across instruments, operators, and staining runs were 1.8%, 2.0%, and 5.1%, respectively. For positive/negative classification between parameters, concordance rates ranged from 94.5% to 99.3% for all cases. Differentially classified cases only occurred around the determined cut point, not over the entire distribution.
  
  
  CONCLUSIONS
  These data demonstrate that AQUA analysis can provide a standardized HER2 immunohistochemistry test that can meet current guidelines by the American Society of Clinical Oncology/College of American Pathologists. The use of AQUA analysis could allow for standardized and objective HER2 testing in clinical laboratories.
authors.affiliation: Department of Development, HistoRx Inc, New Haven, CT 06511, USA. mgustavson@historx.com
authors.forename: Mark D
authors.initials: MD
authors.lastname: Gustavson
authors.forename: Brian
authors.initials: B
authors.lastname: Bourke-Martin
authors.forename: Dylan
authors.initials: D
authors.lastname: Reilly
authors.forename: Melissa
authors.initials: M
authors.lastname: Cregger
authors.forename: Christine
authors.initials: C
authors.lastname: Williams
authors.forename: Jane
authors.initials: J
authors.lastname: Mayotte
authors.forename: Maciej
authors.initials: M
authors.lastname: Zerkowski
authors.forename: Greg
authors.initials: G
authors.lastname: Tedeschi
authors.forename: Robert
authors.initials: R
authors.lastname: Pinard
authors.forename: Jason
authors.initials: J
authors.lastname: Christiansen
mesh_terms: D000465:Algorithms; D014408:Biomarkers, Tumor / Q000378:metabolism; D001943:Breast Neoplasms / Q000378:metabolism* / Q000473:pathology; D018270:Carcinoma, Ductal, Breast / Q000378:metabolism* / Q000473:pathology; D005260:Female; D019084:Fluorescent Antibody Technique, Indirect / Q000379:methods / Q000592:standards*; D017408:Guidelines as Topic; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D008875:Middle Aged; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000378:metabolism*; D015203:Reproducibility of Results; D046888:Tissue Array Analysis
chemical_list: D014408:Biomarkers, Tumor; D018719:Erb-b2 Receptor Tyrosine Kinases; C508053:ERBB2 protein, human