Duplex-specific nuclease and Exo-III enzyme-assisted signal amplification cooperating DNA-templated silver nanoclusters for label-free and sensitive miRNA detection

Abstract

Development of novel miRNA detection strategies plays a crucial role in fundamental research and clinical diagnosis of various diseases, such as infantile pneumonia. We herein develop a rapid and sensitive DNA-templated AgNCs-based miRNA detection approach, pinning the hope on an improved detection sensitivity in an easy-to-operate way. In the method, a hairpin probe is designed to specifically bind with target miRNA, and to initiate the DSN enzyme and Exo-III-assisted dual signal recycles. The resultant guanine-rich DNA sequences after signal amplification turn on the fluorescence of the dark AgNCs by hybridizing with the DNA template of the dark AgNCs. The generated signals are correlated with the amounts of target miRNA in the sensing system. Through a series of experiments, the established approach exhibits a great dynamic range of more than seven orders of magnitude with a low limit of detection of 245 aM, holding great promises for miRNA-related researches and disease diagnosis.Graphical abstract