The development of low-cost, disposable electrochemical
sensors
is an essential step in moving traditionally inaccessible quantitative
diagnostic assays toward the point of need. However, a major remaining
limitation of current technologies is the reliance on standardized
reference electrode materials. Integrating these reference electrodes
considerably restricts the choice of the electrode substrate and drastically
increases the fabrication costs. Herein, we demonstrate that adoption
of two-electrode detection systems can circumvent these limitations
and allow for the development of low-cost, paper-based devices. We
showcase the power of this approach by developing a continuous flow
assay for urinary creatinine enabled by an embedded graphenic two-electrode
detector. The detection system not only simplifies sensor fabrication
and readout hardware but also provides a robust sensing performance
with high detection efficiencies. In addition to enabling high-throughput
analysis of clinical urine samples, our two-electrode sensors provide
unprecedented insights into the fundamental mechanism of the ferricyanide-mediated
creatinine reaction. Finally, we developed a simplified circuitry
to drive the detector. This forms the basis of a smart reader that
guides the user through the measurement process. This study showcases
the potential of affordable capillary-driven cartridges for clinical
analysis within primary care settings.