Improving electrochemical hybridization assays with restriction enzymes

Nucleic acids in blood are early indicators of disease that could be detected by point-of-care biosensors if sufficiently sensitive and facile sensors existed. Electrochemical hybridization assays are sensitive and specific but are limited to very short nucleic acids. We have developed a restriction enzyme-assisted electrochemical hybridization (REH) assay for improved nucleic acid detection. By incorporating target-specific restriction enzymes, we detect long nucleic acids, with performance dependent on the location of the cut site relative to the electrode surface. Thus, we have further established guidelines for REH design to serve as a generalizable platform for robust electrochemical detection of long nucleic acids.