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Optimization of a Multiplex RNA-based Expression Assay Using Breast Cancer Archival Material
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HER2 mRNA transcript quantitation in breast cancer.

K Meehan1, B Clynick2, B Mirzai2,3

  • 1School of Pathology and Laboratory Medicine (M504), University of Western Australia, Crawley, WA, 6009, Australia. katie.meehan@uwa.edu.au.

Clinical & Translational Oncology : Official Publication of the Federation of Spanish Oncology Societies and of the National Cancer Institute of Mexico
|November 13, 2016
PubMed
Summary

Digital droplet PCR (ddPCR) accurately quantifies human epidermal growth factor receptor 2 (HER2) mRNA in breast cancer FFPE tissues. This method shows potential for precise HER2 transcript quantification in clinical settings.

Keywords:
Breast cancerDigital droplet PCRHuman epidermal growth factor receptor

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Area of Science:

  • Oncology
  • Molecular Diagnostics
  • Biotechnology

Background:

  • Accurate human epidermal growth factor receptor 2 (HER2) status is crucial for breast cancer prognosis and treatment selection.
  • Current HER2 assessment relies on protein expression (immunohistochemistry) and gene amplification (in situ hybridisation).
  • Exploring novel quantitative methods for HER2 assessment is essential for improved patient care.

Purpose of the Study:

  • To investigate the utility of quantitative droplet digital PCR (ddPCR) for the detection and absolute quantification of HER2 mRNA.
  • To assess the correlation between HER2 mRNA levels and established HER2 statuses in breast cancer.
  • To evaluate ddPCR's performance on formalin-fixed paraffin-embedded (FFPE) breast cancer specimens.

Main Methods:

  • Digital droplet PCR (ddPCR) was employed to measure HER2 mRNA levels in 178 FFPE breast cancer samples.
  • Samples included HER2-positive, -equivocal, and -negative cases, analyzed from both core biopsies and tissue sections.
  • Standard criteria were used to define HER2 status for comparison with ddPCR results.

Main Results:

  • Significantly higher HER2 mRNA levels were detected by ddPCR in HER2-positive cases compared to equivocal and negative cases (p < 0.025).
  • A cutoff of 490 copies/µl effectively distinguished between HER2-positive and HER2-negative cases.
  • ddPCR results demonstrated consistency between core biopsy and tissue section analyses.

Conclusions:

  • ddPCR is a viable method for quantifying HER2 mRNA transcripts in FFPE breast cancer specimens.
  • The study highlights ddPCR's potential for accurate HER2 transcript quantification in FFPE tissues.
  • Further validation in larger cohorts is necessary to establish a clinically applicable cutoff for ddPCR-based HER2 assessment.