One-pot detection of multiplex prostate cancer miRNAs based on quantum dot-labeled hybridization chain reaction and polydopamine nanoparticle quenching

Abnormal miRNA expression accompanies prostate cancer (PCa) development, making the simultaneous detection of multiple miRNAs a promising strategy for accurate diagnosis. Herein, we developed a novel one-pot method for multiplex miRNAs detection by integrating quantum dot-labeled hybridization chain reaction (HCR) with polydopamine nanoparticles (PDA NPs) quenching. Target miRNA unfold the quantum dot-labeled hairpin probes to initiate HCR for signal amplification. Subsequently, PDA NPs effectively quench unreacted probes to reduce the nonspecific background signal, while the HCR products retain strong fluorescence. This strategy enables the simultaneous detection of multiple miRNAs under a single excitation wavelength, exhibiting picomolar sensitivity. It demonstrates high specificity, capable of discriminating both homologous family members and single-base mismatches of target miRNAs. Furthermore, the method exhibited reliable performance in the analysis of clinical serum samples. This work provides a robust and promising tool for non-invasive miRNA profiling, which could significantly enhance the precise diagnosis of prostate cancer.