Peer-Reviewed Journal Details
Mandatory Fields
Heery B.;Briciu-Burghina C.;Zhang D.;Duffy G.;Brabazon D.;O'Connor N.;Regan F.
ColiSense, today's sample today: A rapid on-site detection of β-d-Glucuronidase activity in surface water as a surrogate for E. coli
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Optional Fields
E. coli Enzyme assay Fluorescence On-site Recreational water β-d-glucuronidase
© 2015 Elsevier B.V. A sensitive field-portable fluorimeter with incubating capability and triplicate sample chambers was designed and built. The system was optimised for the on-site analysis of E. coli in recreational waters using fluorescent based enzyme assays. The target analyte was β-d-Glucuronidase (GUS) which hydrolyses a synthetic substrate 6-Chloro-4-Methyl-Umbelliferyl-β-d-Glucuronide (6-CMUG) to release the fluorescent molecule 6-Chloro-4-Methyl-Umbelliferyl (6-CMU). The system was calibrated with 6-CMU standards. A LOD of 5 nM and a resolution of less than 1 nM was determined while enzyme kinetic tests showed detection of activities below 1 pmol min-1 mL-1 of sample. A field portable sample preparation, enzyme extraction protocol and continuous assay were applied with the system to analyse freshwater and marine samples. Results from a one day field trial are shown which demonstrated the ability of the system to deliver results on-site within a 75 min period.
Grant Details