A Dual-Activated Catalytic Hairpin Assembly Signal Amplification Nanoplatform for High-Sensitivity Detection of Dual-Target MicroRNAs

Early diagnosis of hepatocellular carcinoma is crucial for improving patient prognosis, and microRNA (miRNA) detection via liquid biopsy is important. However, the low abundance, high homology, and complex background interference of miRNAs in biological samples pose serious challenges to the sensitivity and specificity of their detction techniques. This study developed a catalytic hairpin assembly (CHA) signal amplification platform with dual activation of near-infrared light (NIR) and glutathione (GSH), combined with the low background fluorescence properties of upconversion nanoparticles (UCNPs) for the accurate and specific quantification of both miRNA-21 and miRNA-221. Under optimized conditions, the limits of detection for miRNA-21 and miRNA-221 were 0.266 and 0.113 nM, respectively. The recovery rates in the serum spiked experiments ranged from 98.0 to 107.1%, with a low relative standard deviation (<4.84%), suggesting that the method has good accuracy and reliability. The constructed NIR/GSH dual-activation CHA-sensing platform not only offers a novel approach for the determination of low-abundance miRNAs in complex biological samples but also establishes a feasible foundation for the development of advanced theranostic nanoplatforms for early screening and subtyping diagnosis of hepatocellular carcinoma.