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Bacteriophage-enabled Simple Electrochemical Bioassay for the Sensitive Detection of Escherichia Coli in Drinking Water
Wednesday, March 10, 2021: 3:05 PM - 3:25 PM
Speaker(s)
Description
Bacteriophages offer a unique opportunity for highly specific detection of viable bacteria due to their inherent host specificity. Their easy-to-manipulate genome enables rapid, sensitive and simple detection platforms. Here, we engineered the lytic T7 bacteriophage (phage) to overexpress alkaline phosphatase fused to a cellulose-specific carbohydrate-binding module (ALP-CBM). In the bioassay, a sample containing E. coli is infected by the phage in a 30 min incubation period, during which large quantities of ALP-CBM are expressed and freely released into the media. The fusion protein as a direct correlate to the original bacterial concentration, is concentrated out of the media via its CBM function by simple filtration through a cellulose disc, followed by substrate addition and quantification via differential pulse voltammetry using screen-printed carbon electrodes. This extremely simple bioassay resulted in a limit of detection of 103 CFU∙mL-1 within just 2 h. By adding a couple of hours of pre-incubation time for bacterial growth, this sensitivity could be lowered to less than 10 cells per sample. Alternatively, larger volumes can easily be processed. The simplicity of the low-cost bioassay associated with its ruggedness against matrix interferences or reporter concentrations that typically plague antibody-based biosensors and bioassays make it a relevant ,inexpensive and very simple detection system for viable bacteria in complex matrices especially for resource-limited settings. The here presented principle can easily be adapted for the specific detection of other pathogens and paves the way for a new class of biosensors for the identification and quantification of bacteria. Furthermore, multi-analyte detection strategies can easily be realized through the use of various reporter enzymes.
Additional Info
Keywords: Please select up to 4 keywords ONLY:
Enzyme Assays,Food Safety,Genetic Engineering
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