Multiplex Detection of Metastatic Breast Tissue Biomarkers by Fluorescence Spectroscopy

Abstract

Detection of primary lesions prior to the formation of late-stage metastatic tumors is crucial to decreasing mortality rate in breast cancer cases. Research herein contains assays based around three mRNA biomarkers including cytokeratin 19 (CK-19), prolactin inducible protein (PIP), and human epidermal receptor 2 (HER2). These biomarkers are over expressed in breast tumor tissue. Fluorescence molecular beacon detection was used to determine presence of biomarker mRNA. Single MB-mRNA assays provide evidence of ability to detect biomarker mRNA with complementary MBs. The fluorescence detection of biomarkers described herein offers several advantages over current detection methods, providing quantitative accuracy, low limits of detection, and specificity that is expected to result in few false positives and negatives. Our multiplex assay has the additional advantages of efficiency and speed, allowing for higher throughput of samples. Ultimately, multiplex MB-mRNA assays could allow enhanced accuracy, precision, efficiency, and a further understanding of disease progression.