Development of a MicroBio PAD with Enzyme-linked Immunosorbent Assay for the detection of Escherichia coli in water

Elizabeth Chandler Saint Mary's College
Faculty Sponsor(s): Don Paetkau Saint Mary's College
Microbial contamination of water indicates the presence of disease-causing microorganisms, mostly of fecal origin in water, and is associated with gastrointestinal illnesses worldwide. Though several methods of detecting contaminating microbes in water are available, a simple, fast, sensitive, and economic method is still needed. This research intended to develop a prototype of an ELISA-MicroBio PAD, a microbiological paper-based device that utilizes the Enzyme-Linked Immunosorbent Assay (ELISA) to detect Escherichia coli, a common bacterial indicator of fecal contamination in water. A basic ELISA method was adapted on a filter paper and subjected to a series of optimization experiments in order to minimize the time and the reagents while maintaining the sensitivity of detection. A negative control utilizing Enterococcus faecalis was used to compare development of color by Escherichia coli. Optimization experiments included adjusting the filter paper, washing steps, amounts of the washing buffers, and antibodies. On the newly developed prototype, Escherichia coli produced a more rapid and intense color change distinguishable from that produced by the negative control. The prototype was capable of detecting 10^9 CFU/mL using only 5 μL of each reagent. With further optimization and field-testing experiments for validation, this device could be of significant use in detecting microbial contaminants in drinking water.
Biochemistry & Molecular Biology
Poster Presentation

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Irwin Library 3rd Floor