As part of the Food Safety Modernization Act, the FDA has proposed new rules requiring farmers to regularly monitor for “generic” E. coli as an indicator of agricultural water quality. Farmers will need new easy-to-use, rapid, and affordable testing methods to meet these new rules and respond to changes in water quality. This paper describes a phage amplification-based scheme combined with a later flow assay for the detection of generic E. coli. The use of bacteriophage as a detection element has several advantages due to their potential broad host range, short amplification time, and ability to differentiate between viable and non-viable cells. The simplicity, portability, and reliability of paper fluidic devices make them an ideal platform for testing in resource limited settings like those found on-farm. In this study we (i) developed a later flow assay for the detection of T7 bacteriophage; (ii) investigated the impact of low E. coli concentrations and initial T7 inoculum on phage amplification; and ( iii) the subsequent impact on our schemes ability to detect E. coli. Our data indicates that the phage amplification-based lateral flow assay can detect 100 CFU/mL of E. coli following an 8 hour incubation, and 1000 CFU/mL following a 5 hour incubation. The results of this study suggest that, with some modification, a phage amplification-based lateral flow assay for bacterial detection in a format amenable testing in resource-limited setting is feasible, and potentially expandable to other bacterial targets and sample matrices.

Phage amplification-based paper-fluidic device for the detection of generic E. coli

249th National Meeting (2015)

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