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 10 5 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.
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Demonstration of Tryptophan-Like Fluorescence Sensor Concepts for Fecal Exposure Detection in Drinking Water in Remote and Resource Constrained Settings
Low-cost, field-deployable, near-time methods for assessing water quality are not available when and where waterborne infection risks are greatest. We describe the development and testing of a novel device for the measurement of tryptophan-like fluorescence (TLF), making use of recent advances in deep-ultraviolet light emitting diodes (UV-LEDs) and sensitive semiconductor photodiodes and photomultipliers. TLF is an emerging indicator of water quality that is associated with members of the coliform group of bacteria and therefore potential fecal contamination. Following the demonstration of close correlation between TLF and E. coli in model waters and proof of principle with sensitivity of 4 CFU/mL for E. coli, we further developed a two-LED flow-through configuration capable of detecting TLF levels corresponding to “high risk” fecal contamination levels (>10 CFU/100 mL). Findings to date suggest that this device represents a scalable solution for remote monitoring of drinking water supplies to identify high-risk drinking water in near-time. Such information can be immediately actionable to reduce risks.
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- Award ID(s):
- 1738321
- NSF-PAR ID:
- 10229164
- Date Published:
- Journal Name:
- Sustainability
- Volume:
- 12
- Issue:
- 9
- ISSN:
- 2071-1050
- Page Range / eLocation ID:
- 3768
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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- Free Publicly Accessible Full Text
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Accepted Manuscript1.0
- Journal Article:
- https://doi.org/10.3390/su12093768
