There is a growing need for virus-detecting sensors with improved sensitivity and dynamic range, for applications including disease diagnosis, pharmaceutical research, agriculture and homeland security. We report a novel electrochemical biosensing method for improving the sensitivity for detection of the bacteriophage virus MS2, using nanoporous oxirane-derivatized beads. These beads are a commercial polymethyl-metacrylate (PMMA) polymer that has extremely high surface area to volume ratio, making it an ideal platform for surface based sensors. We have developed and evaluated a method for covalent bioconjugation of antibodies and biological support to polymeric beads. The resulting Solid State Kits (SSK) were used to selectively capture enzyme-labeled MS2 viruses from different solutions, enabling detection of a viral concentration of as low as 10 plaque-forming units per milliliter (pfu ml -1 ) by measuring the current (A) from the exposed SSK beads to the enzymatic reaction electrons movement not clear. The kit is connected to a "home made" designed micro-flow system, that exhibits sensitivity and dynamic range similar to the ELISA immuno-liquid array-based assay while outperforming protein micro-array methods.Immuno-Amperometric techniques, using nano-Bio-Polymers Solid Phase Disposable Kit, were used to measure and thus to validate the accuracy of novel technology for virus concentration determination. These work demonstrate the utility of immunoelectrochemical techniques for use in environmental-health quality assurance measurements of viruses.

Abstract:In this work, it is proposed a POCT and innovative method of immunoassay for the detection of C-reactive protein and IgG, using Amperometry coupled to solid state kit, connected to a micro-flow system with comparable sensitivity to a high sensitivity CRP ELISA (hsCRP) and IgG ELISA, with 1-3 min turnaround time to result. Samples of CRP (0 to 250 ng·mL -1 ) andIgG or diluted spiked human serum are injected through a solid state polymeric kit, micro-flow sensor channels of a SWCT SPE nano-modified biosensor. Preparing two kits immuno-assays, in the same micro-column, built on oxirane groups of polymeric bead surface, with biological support to CRP and IgG biomarkers recognition, in a real time scheme, at the end of analyte injection the initial rate of change in current intensity I/A was proportional to CRP respectively IgG concentration, with low detection limit (LOD) of 0.1 ng·mL -1 . It was revealed that CRP/IgG concentrations in serum that might be expected in both normal and pathological conditions can be detected in a real-time-efficient, multi-immunoassay with solid state detection kit technology with determined CRP/IgG concentrations in close agreement with those determined using a commercially available high sensitivity ELISA.