Abstract
Abstract The monitoring of uracil DNA glycosylase (UDG) activity in a simple and sensitive way is very important to clinical diagnosis and biochemical research. In this paper, based on a DNA three junction-mediated recycling signal amplification strategy, we have developed a sensitive and convenient electrochemical approach for monitoring UDG activity and its inhibition. The target UDG specifically recognizes and hydrolyzes uracil bases located in the DNA duplexes, causing the release of the methylene blue-tagged ssDNAs (MB-DNAs) to trigger the disassembly of the three way junction (TWJ) probes and the attachment of the MB-DNAs on the sensor electrode through toehold-mediated strand displacement reaction (TSDR). The disassembled sequences from the TWJ probes further initiate another TSDR to cause cyclic capture of lots of MB-DNAs on the sensing electrode, generating significantly amplified current outputs for sensitively detecting UDG at the low 0.00028 U/mL level. Moreover, such UDG sensing method shows high sensitivity and can be used to screen the inhibition effect of the uracil glycosylase inhibitor to UDG activity, demonstrating the great potential of the developed sensing method for clinical applications.
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