Real Time Diagnostic Point of Care by Amperometric Immuno-Biosensor Kit by Flow Technology

摘要

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 EL1SA 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 immuno-electrochemical techniques for use in environmental-health quality assurance measurements of viruses.