Label-free miRNA fluorescent biosensors based on duplex-specific nucleases and silver nanoclusters

MicroRNAs (miRNAs) are considered reliable biomarkers for a variety of diseases. However, their low abundance in organisms and high sequence similarity of homologous miRNAs make their accurate detection challenging. Here, we constructed a novel fluorescent biosensor for the detection of miRNA-155, a potential biomarker of neuroinflammation, based on duplex-specific nuclease (DSN) assisted amplification and DNA-templated silver nanoclusters (DNA-AgNCs) as fluorescence signal probes. DSN-assisted amplification can transform unstable miRNA into stable DNA and amplify the miRNA signal at the same time. Using DNA-AgNCs as fluorescence signal probes for biosensors can avoid complex labeling processes and reduce costs. The biosensor shows excellent selectivity, reproducibility, a wide linear range (1-600 nM) with a detection limit of 0.86 nM, and potentiality for real sample detection. This work provides a potential universal biosensing platform for miRNA detection.