Simultaneous Detection of Bladder Cancer Exosomal MicroRNAs Based on Inorganic Nanoflare and DNAzyme Walker

Bladder cancer (BC) is one of the most common cancers in the world, with high morbidity and mortality. It is essential to develop a non-invasive, highly accurate, and simple method for BC diagnosis. This work proposed a fluorescent biosensor based on inorganic nanoflares combined with a DNAzyme walker for the simultaneous detection of BC exosomal microRNAs (miRNAs). This biosensor was constructed on the Au nanoparticle (AuNP) modified with the carbon dot (CD)-labeled substrates and DNAzyme strands (AuNP@CDs inorganic nanoflares-DNAzyme, APCD). In the presence of target miRNAs, DNAzyme was activated and then cleaved the CD-labeled substrates and automatically walked along the AuNP, allowing fluorescence recovery. Due to the structure and functional composition, the APCD biosensors demonstrated high sensitivity and specificity, with the reached limit of detection for a single miRNA at the femtomolar level and wide linear range from 50 fM to 10 nM. Furthermore, the simultaneous analysis of BC-related exosomal miR-133b and miR-135b in clinical serum specimens was achieved and consistent with qRT-PCR, suggesting it is a potential method for the diagnosis of BC and other cancers.