FRET-Based Detection of Triple Negative Breast Cancer

Authors

Srishty Sethi, Virginia Commonwealth University

Original Publication Date

2026

Document Type

Video

Comments

Presented in the New Frontiers in Detecting Disease and Managing Pain session.

Abstract

Triple Negative Breast Cancer (TNBC) is the most aggressive breast cancer subtype, characterized by the absence of estrogen, progesterone, and HER2 receptors. Due to its high recurrence rate and resistance to standard hormone therapies, early detection is critical. This presentation describes the development of a novel, non-invasive analytical method using Single-Molecule Fluorescence Resonance Energy Transfer (smFRET) to detect dysregulated microRNA (miRNA) biomarkers directly from body fluids like blood serum.

While miRNAs are ideal biomarkers for early diagnosis, their low physiological concentrations (femtomolar to picomolar) and the fact that single miRNAs are often associated with multiple cancers necessitate a multiplexed detection approach. Traditional methods like RT-PCR require complex RNA extraction and amplification, which can introduce biases. To address these limitations, the researcher leveraged the dynamic conformational switching of DNA four-way (Holliday) junctions.

Key Technological Innovations:

• Four-Way Junction Sensors: The team engineered sensors that remain in a static FRET state in the absence of a target but switch between two distinct FRET states (dynamic signal) upon binding to a specific miRNA.
• Multiplexing via "Backward Assignment": By positioning donor and acceptor fluorophores at varying distances across four different sensors on a single slide, the researchers achieved non-overlapping, distinguishable dynamic signals for four different miRNA targets simultaneously.
• Extraction-Free Detection: Unlike gold-standard methods, this smFRET technology operates directly on serum and plasma samples without requiring RNA extraction, labeling, or amplification.
• High Sensitivity and Specificity: The method demonstrates a limit of detection (LOD) between 10 to 50 femtomolar and shows zero cross-binding in complex biological matrices.

Keywords

Triple-Negative Breast Cancer (TNBC), Single-Molecule FRET (smFRET), MicroRNA (miRNA) biomarkers, Multiplexed detection, Early cancer diagnosis

Rights

Copyright © 2026 Srishty Sethi. All rights reserved.