We have shown that molecular imprinting (MI) technology can be used to produce sensitive, robust, cost-effective biosensing systems with a real-time electrochemical readout that can be utilized for point of care diagnostics. Real time detection of biomarkers is essential when rapid, critical decisions need to be made, such as in situations where public health is threatened. Our biosensor has high sensitivity compared to standard methods like ELISA, and results are obtained within minutes, using inexpensive, accessible potentiometric readout technology. These biosensors utilize surface molecular imprinting of a self-assembling monolayer of hydroxy-terminated alkanethiol chains which form a crystalline ‘lock-and-key’ structure around a target analyte, allowing the sensors to detect and differentiate between bio-macromolecules of similar size and shape with high selectivity and sensitivity. The sensors are extremely versatile and able to detect a diverse range of molecules of varied chemical composition and structure. To fully exploit the sensors’ advantages, especially in remote, economically disadvantaged areas, it is important to quantify their durability and reusability. Biosensor chips were created to test the viability of hemoglobin detection and to evaluate the potential for sensor reusability when washed after detection testing. The successful readsorption of hemoglobin even after washing, accompanied by cyclic voltammetry data indicating the preservation of the SAM, indicate that these biosensors are reusable, significantly augmenting the device’s value. Potential applications include the analysis of complex chemical and biological processes such as stem cell differentiation and on-the-spot detection of diseases such as Zika.