Campus Units

Food Science and Human Nutrition

Document Type

Article

Publication Version

Accepted Manuscript

Publication Date

12-12-2018

Journal or Book Title

Analyst

DOI

10.1039/c8an01850b

Abstract

The monitoring of drinking water for indicators of fecal contamination is crucial for ensuring a safe supply. In this study, a novel electrochemical method was developed for the rapid and sensitive detection of Escherichia coli (E. coli) in drinking water. This strategy is based on the use of engineered bacteriophages (phages) to separate and concentrate target E. coli when conjugated with magnetic beads, and to facilitate the detection by expressing gold binding peptides fused alkaline phosphatase (GBPs-ALP). The fusion protein GBPs-ALP has both the enzymatic activity and the ability to directly bind onto a gold surface. This binding-peptide mediated immobilization method provided a novel and simple approach to immobilize proteins on a solid surface, requiring no post-translational modifications. The concentration of E. coli was determined by measuring the activity of the ALP on gold electrodes electrochemically using linear sweep voltammetry (LSV). This approach was successfully applied in the detection of E. coli in drinking water. We were able to detect 105 CFU mL−1 of E. coli within 4 hours. After 9 hours of preincubation, 1 CFU of E. coli in 100 mL of drinking water was detected with a total assay time of 12 hours. This approach compares favorably to the current EPA method and has the potential to be applied to detect different bacteria in other food matrices.

Comments

This article is published as Wang, D., Hinkley,T, Chen,J., Talbert, J.N., Nugen, S.R., Phage based electrochemical detection of Escherichia coli in drinking water using affinity reporter probes. Analyst, 2018, Doi: 10.1039/C8AN01850B . Posted with permission.

Copyright Owner

Royal Society of Chemistry

Language

en

File Format

application/pdf

Published Version

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