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Updated: Jan 15, 2026
Probe-based Real-time PCR Approaches for Quantitative Measurement of microRNAs
Published on: April 14, 2015
A Pipette-Tip-Based Point-of-Care Test Platform for Detection of MicroRNA at Subfemtomolar Concentrations Using
Mohammad Sadraeian1,2, Jiajia Zhou1,2, Jiayan Liao1
1Institute for Biomedical Materials and Devices (IBMD), Faculty of Science, University of Technology Sydney, Sydney, NSW 2007, Australia.
Abstract:
MicroRNAs are potent early disease biomarkers, yet ultrasensitive assays suitable for point-of-care testing (POCT) remain scarce. We present a pipet-tip platform for quantitative detection of oncogenic miR-27a. This method employs a sandwich hybridization strategy utilizing biotinylated capture oligos on microbeads and upconversion nanoparticle (UCNP)-conjugated oligos as the reporting probe. Highly doped UCNPs were incorporated to enhance the detection sensitivity. The dynamic mobility of the bead-oligo complexes within the pipet tip fluid significantly improves the hybridization efficiency and signal detection specificity. The assay is linear from 128 attomolar (aM) to 2.05 fM, achieves a 158 aM limit of detection (LOD), and yields a 33-fold signal advantage over mismatched strands. Our platform offers a simplified, rapid, and cost-effective solution for detecting low-abundance miRNAs with the reusability of functionalized miRNA-beads. Its scalability and minimal instrumentation requirements make it a strong candidate for advanced molecular diagnostics and POCT applications.
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