Microfluidic Chip for the Electrochemical Detection of MicroRNAs: Methylene Blue Increasing the Specificity of the Biosensor.

MicroRNAs (miRNAs) are biomarkers involved in biological processes that are released by cells and found in biological fluids such as blood. The development of nucleic acid-based biosensors has significantly increased in the past 10 years because the detection of such nucleic acids can easily be applied in the field of early diagnosis. These biosensors need to be sensitive, specific, and fast in order to be effective. This work introduces a newly-built electrochemical biosensor that enables a fast detection in 30 min and, as a result of its integration in microfluidics, presents a limit of detection as low as 1 aM. The litterature concerning the specificity of electrochemical biosensors includes several studies that report one base-mismatch, with the base-mismatch located in the middle of the strand. We report an electrochemical nucleic acid biosensor integrated into a microfluidic chip, allowing for a one-base-mismatch specificity independently from the location of the mismatch in the strand. This specificity was improved using a solution of methylene blue, making it possible to discriminate a partial hybridization from a complete and complementary hybridization.

An Integrated Multiple Electrochemical miRNA Sensing System Embedded into a Microfluidic Chip.

In this article, we present the design, fabrication and characterization of a microfluidic device and a dedicated electronic system to perform 8 multiplexed electrochemical measurements of synthetic miRNA strands, as well as the biochemical protocols developed for the functionalization of the electrodes and the quantification experiments. The outcomes of this work highlight that the parallelization of eight microchannels containing 2-electrode cells driven by the dedicated electronics offers a solution as robust as a conventional 3-electrode cell and commercially available potentiostats. In addition, this solution presents the advantage of simultaneously reduce the microfabrication complexity, as well as offering an integrated; multiplexed and portable system for the quantification of miRNA. The results presented demonstrate that the system shows a linear response on concentrations down to 10-18 mol/L of perfect matched reporter and capture sequences of synthetic miRNA.