The effect of delay in fixation on HER2 expression in invasive carcinoma of the breast assessed with immunohistochemistry and in situ hybridisation

J Clin Pathol. 2014 Jul;67(7):573-5. doi: 10.1136/jclinpath-2013-201978. Epub 2014 Apr 15.

Abstract

Aims: Accurate assessment of HER2 status is essential for selection of patients for HER2-targeted treatment such as trastuzumab. This study investigated the hypothesis that delayed fixation impairs HER2 assessment.

Methods: 9 carcinomas were received fresh, and samples were fixed immediately or put in fixative at time intervals up to 24 h. All carcinomas were scored as 3+ with immunohistochemistry in properly fixed tissue.

Results: 2 of 9 carcinomas (95% CIs 6% to 56%) showed reduced immunohistochemical staining with delays in fixation of 1 and 8 h. One carcinoma showed low-level amplification with fluorescence in situ hybridisation (FISH) when properly fixed and was not amplified after delayed fixation. The other carcinomas were amplified at all time points.

Conclusions: Delayed fixation impaired HER2 protein expression assessed using immunohistochemistry in 22% of 3+ carcinomas. HER2 amplification assessed using FISH may be less affected.

Keywords: Breast; Fixation; Immunohistochemistry; In Situ Hybridisation.

Publication types

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

MeSH terms

  • Biomarkers, Tumor* / analysis
  • Biomarkers, Tumor* / genetics
  • Breast Neoplasms / enzymology*
  • Breast Neoplasms / genetics*
  • Breast Neoplasms / pathology
  • Carcinoma / enzymology*
  • Carcinoma / genetics*
  • Carcinoma / pathology
  • Female
  • Gene Amplification
  • Humans
  • Immunohistochemistry*
  • In Situ Hybridization, Fluorescence*
  • Neoplasm Invasiveness
  • Predictive Value of Tests
  • Prognosis
  • Receptor, ErbB-2* / analysis
  • Receptor, ErbB-2* / genetics
  • Reproducibility of Results
  • Specimen Handling / methods*
  • Time Factors
  • Tissue Fixation*

Substances

  • Biomarkers, Tumor
  • ERBB2 protein, human
  • Receptor, ErbB-2