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Two novel probe-label-free electrochemical aptasensors based on methylene blue (MB)-anchored graphene oxide (GO) amplification were developed for thrombin (TB) and ATP detection, taking advantage of the specific binding affinity of the aptamer towards the target and the different affinities of GO for MB, ssDNA, dsDNA, and G-quadruplex.Scheme 1 Design of aptasensors for TB (A) and ATP (B) detection based on MBanchored GO amplification.

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The focus of this work was on designing a label-free DNA biosensor based on a super-sandwich assay using the electrochemical impedance spectroscopy technique. For this purpose, we designed a signal-up configuration whose linker probes could hybridize with two regions of the target DNA. In this configuration, the presented target DNA would effectively decrease the electron transfer, which would improve the sensitivity of the sensor. Ultimately, we employed gel electrophoresis to further confirm the formation of the proposed super-sandwich structure.