Paper substrates are promising for development of cost-effective and efficient point-of-care biosensors, essential for public healthcare and environmental diagnostics in emergency situations. Most paper-based biosensors rely on the natural capillarity of paper to perform qualitative or semi-quantitative colorimetric detections. To achieve quantification and better sensitivity, technologies combining paper-based substrates and electrical detection are being developed. In this work, we demonstrate the potential of electrical measurements by means of a simple, parallel-plate electrode setup towards the detection of whole-cell bacteria captured in nitrocellulose (NC) membranes. Unlike current electrical sensors, which are mostly integrated, this plug and play system has reusable electrodes and enables simple and fast bacterial detection through impedance measurements. The characterized NC membrane was subjected to (i) a biofunctionalization, (ii) different saline solutions modelling real water samples, and (iii) bacterial suspensions of different concentrations. Bacterial detection was achieved in low conductivity buffers through both resistive and capacitive changes in the sensed medium. To capture Bacillus thuringiensis, the model microorganism used in this work, the endolysin cell-wall binding domain (CBD) of Deep-Blue, a bacteriophage targeting this bacterium, was integrated into the membranes as a recognition bio-interface. This experimental proof-of-concept illustrates the electrical detection of 10⁷ colony-forming units (CFU) mL⁻¹ bacteria in low-salinity buffers within 5 min, using a very simple setup. This offers perspectives for affordable pathogen sensors that can easily be reconfigured for different bacteria. Water quality testing is a particularly interesting application since it requires frequent testing, especially in emergency situations.

Paper substrates have shown a high potential for development of cost-effective and efficient point-of-care biosensors, essential for public healthcare and environmental diagnostics. Most paper-based biosensors rely on qualitative colorimetric detection schemes with high limits of detection. To overcome this limitation, technologies that combine paper-based substrates and electrochemical detection are being developed to allow for quantification and achieve better performances. In this work, we explore the potential of dielectric measurements towards electrical detection of whole-cell bacteria in nitrocellulose membranes, a paper-derivative. Impedance spectroscopy was considered to characterize the membranes with and without Bacillus thuringiensis cells, used as model microorganism. To specifically target this bacterial strain, phage endolysin cell-wall binding domain (CBD) encoded by a bacteriophage targeting B. thuringiensis were prepared and integrated into the membranes as recognition biointerface. The fluid sample containing the bacteria is conducted in the membrane through passive capillarity, and the bacteria are specifically immobilized in the test zone. Resulting changes of the dielectric properties of the membrane are sensed through impedance changes, highlighting the contribution of ions in the bacterial detection mechanism. This experimental proof-of-concept illustrates the electrical detection of 10⁸ CFU/mL bacteria in low-salinity buffers within 5 min.