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Author Spotlight: FISH as a Tool for Precise Gene Amplification Assessment in Cancer Specimens
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Author Spotlight: FISH as a Tool for Precise Gene Amplification Assessment in Cancer Specimens

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Droplet digital polymerase chain reaction detection of HER2 amplification in formalin fixed paraffin embedded breast

Yazhen Zhu1, Dan Lu2, Maruja E Lira3

  • 1Guangdong Provincial Hospital of Traditional Chinese Medicine (GPHTCM), Guangzhou, China.

Experimental and Molecular Pathology
|December 3, 2015
PubMed
Abstract

Insights

Droplet digital PCR (ddPCR) accurately measures Human Epidermal growth factor Receptor 2 (HER2) amplification in breast and gastric cancers. This precise method shows high concordance with standard tests, offering a reliable option for HER2 status determination.

Area of Science:

  • Oncology
  • Molecular Diagnostics
  • Biomarker Analysis

Background:

  • Human Epidermal growth factor Receptor 2 (HER2) gene amplification drives tumorigenesis in various solid tumors.
  • HER2 status is critical for targeted therapy, such as trastuzumab, in HER2-positive metastatic breast and advanced gastric cancers.
  • Accurate HER2 quantification is essential for patient diagnosis and treatment selection.

Purpose of the Study:

  • To evaluate droplet digital PCR (ddPCR) as a robust and accurate method for quantifying HER2 amplification levels.
  • To assess the assay performance of ddPCR in clinical formalin-fixed paraffin-embedded (FFPE) breast and gastric carcinoma samples.
  • To compare ddPCR results with established diagnostic methods like immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH).

Main Methods:

  • ddPCR assays were developed using the HER2 amplified SK-BR-3 cell line.
  • HER2 copy number was quantified relative to the chromosome 17 centromere reference gene (CEP17), yielding a HER2:CEP17 ratio.
  • Genomic DNA from 145 Asian patients' FFPE breast and gastric carcinoma specimens was analyzed using ddPCR, IHC, and/or FISH.

Main Results:

  • ddPCR demonstrated high concordance with FISH and IHC across 145 clinical cases.
  • In breast cancer, ddPCR showed 90.9% sensitivity and 100% specificity compared to FISH and IHC.
  • For gastric cancer specimens with concordant FISH and IHC results, ddPCR achieved 95.5% concordance.

Conclusions:

  • ddPCR offers an automated and precise method for evaluating HER2 amplification levels.
  • The assay's high accuracy and concordance make it a potential alternative for defining HER2 status in clinical settings.
  • ddPCR facilitates the evaluation of HER2 amplification in large sample cohorts, supporting diagnostic workflows.
Keywords:
Breast cancerDroplet digital PCRGastric cancerHER2 amplification

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