Surface Blocking-Based Potentiometric Biosensor for Detection of E. coli ATCC 15597 Using Phage MS2 as a Receptor.

Abstract

Nowadays, using a potentiometric ion sensor to achieve detection of biological analytes is still a big challenge, since these analytes usually do not yield a measurable ion signal. To address this challenge, a simple and robust potentiometric sensing protocol based on a delayed Nernstian response is proposed for the label-free detection of biological analytes. The proposed sensor platform is composed of two layers: the surface recognition layer and the indicator-ion-selective electrode (ISE) membrane layer. It is based on a surface blocking mechanism in which the surface recognition interactions between the surface recognition element and the target can impede the diffusion of the indicator ion from the aqueous phase to the sensing membrane phase to reach the final Nernstian-response equilibrium, thus resulting in a delayed potential change. Such a potential change could be used to measure the concentration of a biological target in the sample. Thus, a sensing system was designed by using phage MS2, its host bacterium Escherichia coli ATCC 15597(abbreviated as E. coli H), and a solid-contact butyrylcholine ISE as a surface recognition element, a target, and an indicator electrode, respectively. This new concept offers a simple, sensitive, and extremely selective potentiometric method for detection of E. coli H with a detection limit of 100 CFU mL-1. It can be expected that the present approach may pave the way to using ISEs to detect various important nonionic biological targets in clinical and environmental applications.