Target-dependent dual strand extension recycling amplifications for non-label and ultrasensitive sensing of serum microRNA

Abstract

MicroRNAs (miRNAs) are important biomarkers because their abnormal expressions or mutations can usually indicate the development of cancers or other diseases. However, their high family sequence homology and low abundance pose a major challenge for the analysis of miRNAs. In the present work, we developed a dual strand extension recycling signal amplification strategy for non-label detection of miRNA-155 with high sensitivity on the basis of DNA polymerase and the G-quadruplex/thioflavin T (ThT) complexes. The specific miRNA-155 target sequences could initiate two strand extension-based independent recycling cycles under the function of the DNA polymerase, resulting in the production of many active G-quadruplex segments. The dye of ThT further bound the G-quadruplexes to induce greatly enhanced fluorescence for ultra-sensitively detecting miRNA-155 sequences at the low concentration of 35 fM in the dynamic range from 0.1 pM to 100 pM. The proposed detection strategy used the completely unmodified and synthetic DNA as probes and could be performed in homogeneous solutions. This method with a high selectivity could also be used for diluted serum samples. The demonstration of our new method for miRNA detection thus offers it with high potentials for miRNA biomarker analyses with the purpose of clinical diagnostics and biomedical applications.