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Building Up a High-throughput Screening Platform to Assess the Heterogeneity of HER2 Gene Amplification in Breast Cancers
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Building Up a High-throughput Screening Platform to Assess the Heterogeneity of HER2 Gene Amplification in Breast Cancers

Published on: December 5, 2017

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Evaluation of HER2 Gene Status in Breast Cancer Samples with Indeterminate Fluorescence in Situ Hybridization by

Vladimira Koudelakova1, Jitka Berkovcova1, Radek Trojanec1

  • 1Institute of Molecular and Translational Medicine, Faculty of Medicine and Dentistry, Palacky University and University Hospital in Olomouc, Olomouc, Czech Republic.

The Journal of Molecular Diagnostics : JMD
|May 10, 2015
PubMed

Insights

A new quantitative real-time PCR method accurately determines HER2 gene amplification in breast cancer. This DNA-based test offers high sensitivity and specificity, especially when FISH assays fail, improving HER2 testing reliability.

Area of Science:

  • Oncology
  • Molecular Diagnostics
  • Biotechnology

Background:

  • HER2 (ERBB2) amplification/overexpression is crucial for breast cancer therapy.
  • Current diagnostic methods like in situ hybridization (ISH) and immunohistochemistry (IHC) have limitations.
  • ISH assays fail in ~5% of breast cancers, leading to potential diagnostic errors with IHC alone.

Purpose of the Study:

  • To develop and validate a quantitative real-time PCR (qPCR) method for determining HER2 DNA copy number.
  • To assess the performance of the qPCR method using formalin-fixed, paraffin-embedded (FFPE) breast cancer tissues.
  • To provide a reliable alternative diagnostic tool when standard HER2 testing methods are inconclusive.

Main Methods:

  • Developed a quantitative real-time PCR assay for DNA extracted from FFPE breast cancer samples.
  • Determined the limit of detection using breast cancer cell lines.
  • Validated the method with 223 patient samples, comparing results against fluorescent ISH (FISH) and IHC data.

Main Results:

  • The qPCR method demonstrated high sensitivity (94.2%-95.1%) and specificity (99.1%-100%) compared to FISH/IHC.
  • Overall concordance between qPCR and FISH/IHC was high (97.6%).
  • Successfully determined HER2 copy number in 69.2% of samples with indeterminate FISH results.

Conclusions:

  • The developed DNA-based qPCR method is a specific and sensitive tool for determining HER2 copy number.
  • This qPCR assay effectively complements IHC tests and provides reliable results when FISH assays fail.
  • The method improves diagnostic accuracy for HER2 status in breast cancer patients, aiding therapeutic decisions.

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