Analytical Validation of a Novel 6-Gene Signature for Prediction of Distant Recurrence in Estrogen Receptor-Positive, HER2-Negative, Early-Stage Breast Cancer
Loughman, Tony; Barron, Stephen; Wang, Chan-Ju Angel; Dynoodt, Peter; Fender, Bozena; Lopez-Ruiz, Cesar; Stapleton, Sharon; Fabre, Aurelie; Quinn, Cecily; Nodin, Bjorn; Jirström, Karin; Razmara, Fatemeh; O'Grady, Anthony; Baird, Anne-Marie; Gray, Steven G; Freixo, Ana; Moelans, Cathy B; van Diest, Paul J; Duffy, Michael J; O'Leary, Desmond; Crown, John; Bracken, Adrian P; Gallagher, William M
(2022) Clinical chemistry, volume 68, issue 6, pp. 837 - 847
(Article)
Abstract
BACKGROUND: OncoMasTR is a recently developed multigene prognostic test for early-stage breast cancer. The test has been developed in a kit-based format for decentralized deployment in molecular pathology laboratories. The analytical performance characteristics of the OncoMasTR test are described in this study. METHODS: Expression levels of 6 genes were measured
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by 1-step reverse transcription-quantitative PCR on RNA samples prepared from formalin-fixed, paraffin-embedded (FFPE) breast tumor specimens. Assay precision, reproducibility, input range, and interference were determined using FFPE-derived RNA samples representative of low and high prognostic risk scores. A pooled RNA sample derived from 6 FFPE breast tumor specimens was used to establish the linear range, limit of detection, and amplification efficiency of the individual gene expression assays. RESULTS: The overall precision of the OncoMasTR test was high with an SD of 0.16, which represents less than 2% of the 10-unit risk score range. Test results were reproducible across 4 testing sites, with correlation coefficients of 0.94 to 0.96 for the continuous risk score and concordance of 86% to 96% in low-/high-risk sample classification. Consistent risk scores were obtained across a > 100-fold RNA input range. Individual gene expression assays were linear up to quantification cycle values of 36.0 to 36.9, with amplification efficiencies of 80% to 102%. Test results were not influenced by agents used during RNA isolation, by low levels of copurified genomic DNA, or by moderate levels of copurified adjacent nontumor tissue. CONCLUSION: The OncoMasTR prognostic test displays robust analytical performance that is suitable for deployment by local pathology laboratories for decentralized use.
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Keywords: RT-qPCR, breast cancer, gene expression, prognostic biomarker, Biochemistry, medical, Clinical Biochemistry, Journal Article
ISSN: 0009-9147
Publisher: American Association for Clinical Chemistry Inc.
Note: Publisher Copyright: © 2022 American Association for Clinical Chemistry.
(Peer reviewed)