HER-2 DNA quantification of paraffin-embedded breast carcinomas with LightCycler real-time PCR in comparison to immunohistochemistry and chromogenic in situ hybridization

Clin Biochem. 2006 Sep;39(9):942-6. doi: 10.1016/j.clinbiochem.2006.06.001. Epub 2006 Jul 13.


Objectives: To compare the detection of HER-2 status by real-time PCR, on paraffin-embedded breast carcinomas, in respect to immunohistochemistry (IHC) and chromogenic in situ hybridization (CISH).

Design and methods: Paraffin-embedded breast carcinomas collected from 85 patients diagnosed with early stage breast cancer were analyzed for HER-2 gene amplification by real-time PCR and CISH, as well as for HER-2 protein expression by IHC.

Results: HER-2 gene amplification was observed in 19 (22.4%) of 85 breast cancer patients by real-time PCR and in 19 (22.4%) of 85 patients by CISH. Strong (3+) HER-2 protein over-expression was observed in 13 (15.3%) out of 85 patients. Moreover, there were 4 out of 85 (4.7%) patients that had moderate (2+) HER-2 protein over-expression, while 68 out of 85 (80%) patients had no HER-2 protein over-expression by IHC. There were strong concordance rates between real-time PCR and IHC (79/85, 92.9%, p<0.0001) and real-time PCR and CISH (77/85, 90.6%, p<0.0001). The concordance rate between the three methods was 90.6% (p<0.0001).

Conclusions: Our data show that the results obtained for amplification of HER-2 by real-time PCR on the LightCycler are comparable to those obtained by IHC and CISH.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Breast Neoplasms / metabolism*
  • Carcinoma / metabolism*
  • Chromogenic Compounds / chemistry
  • DNA / analysis
  • DNA, Neoplasm / analysis*
  • Gene Amplification
  • Gene Dosage
  • Genes, erbB-2*
  • Humans
  • Immunohistochemistry / methods
  • In Situ Hybridization / methods
  • Paraffin Embedding
  • Polymerase Chain Reaction / methods*
  • Receptor, ErbB-2 / genetics*
  • Receptor, ErbB-2 / metabolism


  • Chromogenic Compounds
  • DNA, Neoplasm
  • DNA
  • Receptor, ErbB-2