PMID- 16224218
OWN - NLM
STAT- MEDLINE
DCOM- 20060131
LR  - 20191210
IS  - 0147-5185 (Print)
IS  - 0147-5185 (Linking)
VI  - 29
IP  - 11
DP  - 2005 Nov
TI  - Analytical validation and interobserver reproducibility of EnzMet GenePro: a 
      second-generation bright-field metallography assay for concomitant detection of 
      HER2 gene status and protein expression in invasive carcinoma of the breast.
PG  - 1505-11
AB  - Fluorescence in situ hybridization (FISH) has both excellent sensitivity and 
      specificity in detecting HER2 gene amplification in invasive breast carcinoma. 
      FISH has not been widely implemented in clinical practice because of reagent 
      costs and the special instrumentation and expertise required to perform and 
      integrate the assay. Immunohistochemistry (IHC) for HER2 protein is widely used, 
      but false-positive and false-negative results are problematic. We developed a 
      bright-field assay to visualize HER2 gene amplification and concomitant HER2 
      protein expression (EnzMet GenePro). This assay detects HER2 gene amplification 
      via deposition of metallic silver by enzyme metallographytrade mark (EnzMettrade 
      mark, Nanoprobes, Yaphank, NY) combined with HER2 protein detection by IHC using 
      alkaline phosphatase and fast red K substrate visualization (CB11;Ventana, 
      Tucson, AZ). The assay was performed on 94 invasive breast carcinomas, for which 
      FISH (PathVysiontrade mark, Vysis, Downer's Grove, IL), conventional IHC (CB11), 
      and enzyme metallography (EnzMettrade mark) results were known. The EnzMettrade 
      mark component of the assay was scored as either HER2 gene amplified, polysomic, 
      or nonamplified. The IHC component was scored using the conventional FDA scale of 
      0 to 3+. Concordance of the EnzMet component of the assay versus FISH was 
      assessed and showed an excellent correlation (Pearson coefficient of 0.95; P < 
      0.001). The combination of gene and protein detection (EnzMet GenePro) displayed 
      a specificity of 100% and an accuracy of 92.6% (95% confidence interval 
      85.3-97.0), facilitated recognition of gene/protein discordances, and allowed for 
      efficient interpretation of the slide by conventional light microscopy. The 
      interobserver kappa for each component was excellent (IHC, kappa = 0.94; and 
      EnzMettrade mark, kappa = 0.96). EnzMet is the first bright-field ISH assay in 
      our experience that routinely and nonambiguously detects endogenous HER2 signals, 
      essential for a reliable clinical HER2 assay, and in combination with HER2 
      protein enables improved diagnosis in borderline cases.
FAU - Downs-Kelly, Erinn
AU  - Downs-Kelly E
AD  - Department of Anatomical and Clinical Pathology, Cleveland Clinic Foundation and 
      the Cleveland Clinic Lerner College of Medicine, Case Western Reserve University, 
      Cleveland, OH, USA.
FAU - Pettay, James
AU  - Pettay J
FAU - Hicks, David
AU  - Hicks D
FAU - Skacel, Marek
AU  - Skacel M
FAU - Yoder, Brian
AU  - Yoder B
FAU - Rybicki, Lisa
AU  - Rybicki L
FAU - Myles, Jonathan
AU  - Myles J
FAU - Sreenan, Joseph
AU  - Sreenan J
FAU - Roche, Patrick
AU  - Roche P
FAU - Powell, Richard
AU  - Powell R
FAU - Hainfeld, James
AU  - Hainfeld J
FAU - Grogan, Thomas
AU  - Grogan T
FAU - Tubbs, Raymond
AU  - Tubbs R
LA  - eng
GR  - 1R43 GM064257-01/GM/NIGMS NIH HHS/United States
GR  - 2R42CA83618-02/CA/NCI NIH HHS/United States
GR  - 5R42 CA083618-03/CA/NCI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
PT  - Validation Study
PL  - United States
TA  - Am J Surg Pathol
JT  - The American journal of surgical pathology
JID - 7707904
RN  - EC 2.7.10.1 (Receptor, ErbB-2)
SB  - IM
MH  - Adenocarcinoma/diagnosis/*genetics/immunology
MH  - Breast Neoplasms/diagnosis/*genetics/immunology
MH  - Female
MH  - Genes, erbB-2/*immunology
MH  - Humans
MH  - Immunohistochemistry/*methods
MH  - Observer Variation
MH  - Receptor, ErbB-2/*immunology
MH  - Reproducibility of Results
EDAT- 2005/10/15 09:00
MHDA- 2006/02/01 09:00
CRDT- 2005/10/15 09:00
PHST- 2005/10/15 09:00 [pubmed]
PHST- 2006/02/01 09:00 [medline]
PHST- 2005/10/15 09:00 [entrez]
AID - 00000478-200511000-00012 [pii]
AID - 10.1097/01.pas.0000172294.67409.4f [doi]
PST - ppublish
SO  - Am J Surg Pathol. 2005 Nov;29(11):1505-11. doi: 
      10.1097/01.pas.0000172294.67409.4f.