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Updated: Feb 23, 2026
Optimization of a Multiplex RNA-based Expression Assay Using Breast Cancer Archival Material
Published on: August 1, 2018
Analytical validation of a 12-gene molecular test for the prediction of distant recurrence in breast cancer
M Bryan Warf1,1, Saradha Rajamani2,2, Kristin Krappmann3,3
1Myriad Genetic Laboratories, Inc., Salt Lake City, UT 84108, USA.
Aim:
To validate the analytical performance of a 12-gene molecular assay that predicts distant recurrence for early-stage ER+/HER2- invasive breast cancer as run within a central reference laboratory.
Materials & Methods:
Formalin-fixed paraffin-embedded breast resections were evaluated by quantitative reverse transcription polymerase chain reaction for the expression of eight target genes, three housekeeper genes and one control gene to assess for DNA contamination.
Results:
The assay results were highly correlated with a validated reference laboratory. The assay had a broad linear range for input RNA, with similar amplicon efficiencies for target and housekeeper genes. The assay test was highly reproducible, with comparable inter- and intrabatch precision to the reference laboratory.
Conclusion:
These studies demonstrate that the 12-gene molecular assay is highly robust and accurate.
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