| 초록 |
Herein, a novel enzyme-free and label-free microRNA detection method based on mass spectrometry was developed utilizing catalytic hairpin assembly and DNA-cleaving DNAzyme. In this strategy, hairpin probe 1 (H1) and 2 (H2), respectively composed of DNAzyme sequence and substrate sequence, were used to recognize the target microRNA and amplify the mass peak signal. Target was first hybridized with the toehold region of the H1 and opened H1 was then hybridized with the toehold region of the H2, which led to the formation of DNAzyme-substrate complex and consequently released target. The complex was cleaved into two fragments in the presence of Zn2+ ion, which were then subjected to MALDI-TOF mass spectrometry analysis. Since the target could be continuously recycled to open unreacted H1 probes, a single target could produce numerous mass markers. Thus, for early clinical diagnosis, this new strategy could serve as a promising platform for sensitive detection of the microRNA and further be utilized for multiplex detection of the various microRNAs in a single tube by modulating the sequence of hairpin probes and mass markers. |
