title: Human epidermal growth factor receptor 2 assessment in a case-control study: comparison of fluorescence in situ hybridization and quantitative reverse transcription polymerase chain reaction performed by central laboratories.
abstract: OBJECTIVE
  The optimal method to assess human epidermal growth factor receptor 2 (HER2) status remains highly controversial. Before reporting patient HER2 results, American Society of Clinical Oncology (ASCO)/College of American Pathologists (CAP) guidelines mandate that laboratories demonstrate ≥ 95% concordance to another approved laboratory or methodology. Here, we compare central laboratory HER2 assessed by fluorescence in situ hybridization (FISH) and quantitative reverse transcriptase polymerase chain reaction (RT-PCR) using Oncotype DX in lymph node-negative, chemotherapy-untreated patients from a large Kaiser Permanente case-control study.
  
  
  METHODS
  Breast cancer specimens from the Kaiser-Genomic Health study were examined. Central FISH assessment of HER2 amplification and polysomy 17 was conducted by PhenoPath Laboratories (ratios > 2.2, 1.8 to 2.2, and < 1.8 define HER2 positive, HER2 equivocal, and HER2 negative, respectively). HER2 expression by RT-PCR was conducted using Oncotype DX by Genomic Health (normalized expression units ≥ 11.5, 10.7 to < 11.5, and < 10.7 define HER2 positive, HER2 equivocal, and HER2 negative, respectively). Concordance analyses followed ASCO/CAP guidelines.
  
  
  RESULTS
  HER2 concordance by central FISH and central RT-PCR was 97% (95% CI, 96% to 99%). Twelve percent (67 of 568 patients) and 11% (60 of 568 patients) of patients were HER2 positive by RT-PCR and FISH, respectively. HER2-positive patients had increased odds of dying from breast cancer compared with HER2-negative patients. Polysomy 17 was demonstrated in 12.5% of all patients and 33% of FISH-positive patients. Nineteen of 20 FISH-positive patients with polysomy 17 were also RT-PCR HER2 positive. Although not statistically significantly different, HER2-positive/polysomy 17 patients tended to have the worst prognosis, followed by HER2-positive/eusomic, HER2-negative/polysomy 17, and HER2-negative/eusomic patients.
  
  
  CONCLUSIONS
  There is a high degree of concordance between central FISH and quantitative RT-PCR using Oncotype DX for HER2 status, and the assay warrants additional study in a trastuzumab-treated population.
authors.affiliation: University of California, San Francisco, 1600 Divisadero St, Rm R200, San Francisco, CA 94063, USA. rbaehner@genomichealth.com
authors.forename: Frederick L
authors.initials: FL
authors.lastname: Baehner
authors.forename: Ninah
authors.initials: N
authors.lastname: Achacoso
authors.forename: Tara
authors.initials: T
authors.lastname: Maddala
authors.forename: Steve
authors.initials: S
authors.lastname: Shak
authors.forename: Charles P
authors.initials: CP
authors.lastname: Quesenberry
authors.forename: Lynn C
authors.initials: LC
authors.lastname: Goldstein
authors.forename: Allen M
authors.initials: AM
authors.lastname: Gown
authors.forename: Laurel A
authors.initials: LA
authors.lastname: Habel
mesh_terms: D000328:Adult; D014408:Biomarkers, Tumor / Q000378:metabolism; D001943:Breast Neoplasms / Q000175:diagnosis* / Q000235:genetics / Q000378:metabolism / Q000401:mortality; D002140:California / Q000453:epidemiology; D016022:Case-Control Studies; D002886:Chromosomes, Human, Pair 17; D005188:False Negative Reactions; D005189:False Positive Reactions; D005260:Female; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D007753:Laboratories; D016015:Logistic Models; D008875:Middle Aged; D010338:Pathology, Clinical; D011379:Prognosis; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000378:metabolism*; D012042:Registries; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D014408:Biomarkers, Tumor; D018719:Erb-b2 Receptor Tyrosine Kinases