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Updated: Feb 6, 2026
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Optimization of a Multiplex RNA-based Expression Assay Using Breast Cancer Archival Material
Published on: August 1, 2018
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Optimization of a Multiplex RNA-based Expression Assay Using Breast Cancer Archival Material
Shawn Baldacchino1, Christian Saliba2, Jeanesse Scerri3
1Department of Pathology, Faculty of Medicine & Surgery, University of Malta.
Journal of Visualized Experiments : Jove
|August 21, 2018
Summary
This study presents a novel RNA quantification method for archival cancer tissues, enabling accurate breast cancer subtyping and biomarker measurement. The technique enhances diagnostic accuracy and supports oncology research using valuable historical patient samples.
Area of Science:
- Oncology
- Molecular Diagnostics
- Pathology
Background:
- Archival tissue degradation and limited fresh samples hinder cancer diagnostics.
- Existing gene expression tests require fresh tissue or are less accurate with aged samples.
- Need for robust, workflow-compatible diagnostic tests for archival material.
Purpose of the Study:
- To optimize a branched-DNA (bDNA) assay for accurate RNA quantification in archival cancer tissues.
- To develop a multiplex method for classifying breast cancer molecular subtypes.
- To enable biomarker measurement in archival and liquid biopsy samples for improved patient management.
Main Methods:
- Utilized Invitrogen™ QuantiGene™ Plex Assay with branched-DNA (bDNA) technology.
- Employed Luminex xMAP® magnetic beads for multiplex RNA quantification.
- Integrated laser microdissection for analyzing heterogeneous tumors from H&E stained sections.
Main Results:
- Successfully quantified RNA in archival material, overcoming degradation issues.
- Developed a novel, rapid, and multiplex assay for accurate breast cancer subtyping.
- Demonstrated potential for biomarker measurement in archival and liquid biopsy samples.
Conclusions:
- The optimized bDNA assay provides accurate gene expression profiling from archival tissue.
- This method offers a subjective-free alternative to imaging for cancer classification.
- The assay has broad applications in diagnostics, liquid biopsies, and retrospective oncology research.
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