title: Her-2/neu analysis in archival tissue samples of human breast cancer: comparison of immunohistochemistry and fluorescence in situ hybridization.
abstract: OBJECTIVE
  The objective of our study was to compare the methods used in the literature to analyze HER-2/neu status on archival breast cancer tissue. Therefore, a series of antibodies was evaluated to assess their immunohistochemical (IHC) sensitivity in correlation to gene amplification determined by fluorescence in situ hybridization (FISH).
  
  
  METHODS
  HER-2/neu overexpression was studied on paraffin sections of 85 invasive breast cancers using a panel of five monoclonal (9G6, 3B5, CB11, TAB250, GSF-HER2) and two polyclonal antibodies (A8010, A0485) in addition to the HercepTest (DAKO, Glostrup, Denmark). HER-2/neu gene amplification was determined by FISH using a dual-color probe (PathVysion; Vysis, Stuttgart-Fasanenhof, Germany).
  
  
  RESULTS
  HER-2/neu overexpression was demonstrated in 26% (9G6, TAB250, GSF-HER2), 27% (3B5, CB11), 33% (A8010) and 42% (A0485, HercepTest) of the tumors. FISH on paraffin sections identified gene amplification in 28% of the tumors. Strongly positive IHC results (3+) were always associated with gene amplification. Among the 16 tumors presented with weakly positive IHC results (2+) using the HercepTest, 12 (75%) lacked gene amplification.
  
  
  CONCLUSIONS
  The comparison of IHC and FISH demonstrated an excellent correlation of high-level HER-2/neu overexpression (3+) with gene amplification; ie, FISH does not provide further information in these tumors. However, weakly positive IHC results (2+) obtained with the HercepTest share only a minor association with gene amplification.
authors.affiliation: Pathologisches Institut and Klinik für Frauenheilkunde und Geburtshilfe Klinikum-GroBhadern, Ludwig-Maximilians-Universität, München, Germany. a.lebeau@lrz.uni-muenchen.de
authors.forename: A
authors.initials: A
authors.lastname: Lebeau
authors.forename: D
authors.initials: D
authors.lastname: Deimling
authors.forename: C
authors.initials: C
authors.lastname: Kaltz
authors.forename: A
authors.initials: A
authors.lastname: Sendelhofert
authors.forename: A
authors.initials: A
authors.lastname: Iff
authors.forename: B
authors.initials: B
authors.lastname: Luthardt
authors.forename: M
authors.initials: M
authors.lastname: Untch
authors.forename: U
authors.initials: U
authors.lastname: Löhrs
mesh_terms: D000911:Antibodies, Monoclonal; D001943:Breast Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D005784:Gene Amplification; D018734:Genes, erbB-2; D006801:Humans; D007150:Immunohistochemistry; D017404:In Situ Hybridization, Fluorescence; D021141:Nucleic Acid Amplification Techniques; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000276:immunology / Q000378:metabolism*
chemical_list: D000911:Antibodies, Monoclonal; D018719:Erb-b2 Receptor Tyrosine Kinases