Electrokinetically Driven On-Fiber Active Lateral Flow Assay for MicroRNA Detection
Jawairia Umar Khan1, Gungun Lin1, Mohammad Sadraeian1
1Institute for Biomedical Materials and Devices (IBMD), School of Mathematical and Physical Sciences, Faculty of Science, University of Technology Sydney, Sydney, NSW 2007, Australia.
Analytical Chemistry
|July 7, 2025
Summary
This study introduces an active lateral flow assay (ALFA) for rapid, sensitive detection of the tumor biomarker miRNA-27a using minimal sample volume. ALFA offers a promising advancement for point-of-care testing (POCT) in oncology.
Area of Science:
- Biomarker Discovery
- Molecular Diagnostics
- Point-of-Care Testing
Background:
- MicroRNAs are crucial for early disease detection, especially in oncology.
- Low microRNA concentrations in biospecimens challenge current detection methods, necessitating large volumes or amplification.
- Existing lateral flow assays (LFAs) lack the sensitivity and specificity required for point-of-care testing (POCT).
Purpose of the Study:
- To develop a rapid and sensitive method for detecting miRNA-27a, a key tumor biomarker.
- To overcome the limitations of conventional LFAs by integrating electrophoretic separation.
- To establish a new platform for next-generation POCT with reduced sample requirements.
Main Methods:
- Developed an active lateral flow assay (ALFA) integrating on-fiber electrophoretic separations.
- Utilized upconversion nanoparticles with near-infrared excitation for sensitive detection.
- Optimized electrokinetic cleaning and surface pretreatments for enhanced sensitivity.
Main Results:
- Achieved sensitive detection of miRNA-27a from 1 fM to 10 pM, with a limit as low as 50 attomolar.
- Demonstrated effective separation of target miRNA from complex biomolecules and nonspecific probes.
- Reduced sample volume requirement by 10-fold compared to conventional LFAs.
Conclusions:
- ALFA offers a novel approach for rapid, sensitive, and low-volume microRNA detection.
- The developed system is suitable for next-generation POCT applications in oncology.
- ALFA's cost-effectiveness and scalability support potential industry adoption.
