Abstract

In the developing world, the identification of clean, potable water continues to pose a pervasive challenge, and waterborne diseases due to fecal contamination of water supplies significantly threaten public health. The ability to efficiently monitor local water supplies is key to water safety, yet no low-cost, reliable method exists to detect contamination quickly. We developed an in vitro assay utilizing an odorant-binding protein (OBP), AgamOBP1, from the mosquito, Anopheles gambiae, to test for the presence of a characteristic metabolite, indole, from harmful coliform bacteria. We demonstrated that recombinantly expressed AgamOBP1 binds indole with high sensitivity. Our proof-of-concept assay is fluorescence-based and demonstrates the usefulness of insect OBPs as detector elements in novel biosensors that rapidly detect the presence of bacterial metabolic markers, and thus of coliform bacteria. We further demonstrated that rAgamOBP1 is suitable for use in portable, inexpensive “dipstick” biosensors that improve upon lateral flow technology since insect OBPs are robust, easily obtainable via recombinant expression, and resist detector “fouling.” Moreover, due to their wide diversity and ligand selectivity, insect chemosensory proteins have other biosensor applications for various analytes. The techniques presented here therefore represent platform technologies applicable to various future devices.

Highlights

  • Ensuring water supplies are safe is crucial to public health

  • Water supply safety concerns are not limited to potable water or even water that is intended for domestic use; irrigation water and water sources not intended for direct human use or consumption must meet basic safety and hygiene requirements in order to ensure public safety and prevent disease transmission from water-borne microbes

  • AgamOBP1 binds analytes associated with coliform bacteria and does so with high specificity and sensitivity [18,34], allowing the rapid detection of low level E. coli contamination in water supplies

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Summary

Introduction

Ensuring water supplies are safe is crucial to public health. Water supply safety concerns are not limited to potable water or even water that is intended for domestic use; irrigation water and water sources not intended for direct human use or consumption must meet basic safety and hygiene requirements in order to ensure public safety and prevent disease transmission from water-borne microbes. Protection Agency for detecting E. coli contamination require at least 24 h [3]. An interesting characteristic shared by many current contamination detection methods is the essentially retroactive or ex post facto nature of their results; since point source contamination of water supplies is sometimes swift, a quick, field-deployable device could address water safety concerns on a more immediate basis

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