Hybridization for human epidermal growth factor receptor 2 testing in gastric carcinoma: a comparison of fluorescence in-situ hybridization with a novel fully automated dual-colour silver in-situ hybridization method

Histopathology. 2011 Jul;59(1):8-17. doi: 10.1111/j.1365-2559.2011.03894.x.

Abstract

Aims: Amplification of the human epidermal growth factor receptor 2 (HER2) gene has been reported in gastric carcinoma (GC). Accordingly, trastuzumab plus chemotherapy has recently become the new standard treatment for HER2-positive advanced GCs. The aim was to compare the alleged gold standard for hybridization [fluorescence in-situ hybridization (FISH)] with a novel, fully automated brightfield dual-colour silver-enhanced in-situ hybridization (SISH) method.

Methods and results: The studies series was comprised of 166 GC samples. Additionally, tumours with discordant results obtained by FISH and SISH were analysed by real-time quantitative polymerase chain reaction (PCR) with the LightMix kit HER-2/neu. Of the samples, 17.5% and 21% were amplified by FISH and SISH, respectively. Heterogeneity was identified in up to 52% of cases. In 96.4% of cases, FISH showed the same results as SISH. All six discordant cases were positive by SISH and negative by FISH. On review of the FISH slides, all contradictory cases were polysomic and were confirmed to be negative for amplification by real-time PCR. Interestingly, all ratios in this latter group were between 2.06 and 2.50, so setting the cut-off for amplification at ≥3 resulted in perfect concordance.

Conclusions: Dual-colour SISH represents a novel method for the determination of HER2 status in GC.

Publication types

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

MeSH terms

  • Adenocarcinoma / genetics
  • Automation, Laboratory / methods
  • Gene Amplification
  • Gene Dosage
  • Genes, erbB-2*
  • Humans
  • In Situ Hybridization / methods*
  • In Situ Hybridization / standards
  • In Situ Hybridization, Fluorescence / methods*
  • In Situ Hybridization, Fluorescence / standards
  • Real-Time Polymerase Chain Reaction
  • Silver
  • Stomach Neoplasms / genetics*

Substances

  • Silver