DNAzyme Crosslinked Polyacrylamide Hydrogels for the Colorimetric Detection of E. coli

Abstract

Escherichia coli (E. coli) is a gram-negative bacteria found in the intestinal system of humans that can also contaminate food, drinking water, as well as lakes and rivers. While not all strains are pathogenic, some including O157:H7 can cause severe illness. Conventional methods of detecting E. coli contamination in water samples often have limitations for on-site testing applications, which can include their slow detection time or need for expensive laboratory equipment. While several fluorescent biosensors for the detection of E. coli have been developed in the Didar lab, there is increased interest in colourimetric biosensors whose signal can be interpreted with the naked eye. This thesis will describe the development and performance of a hydrogel biosensor, that is made of polyacrylamide chains crosslinked by an E. coli detecting Deoxyribozyme (DNAzyme) and its substrate. In the presence of E. coli, the DNAzyme cleaves its substrate and crosslinking breaks down, resulting in the visible dissolution of the hydrogel. Paired with the use of bacteriophage induced cell lysis to amplify the target protein, detection sensitivity to the order of 10^1 CFU/mL has been achieved using this platform with an incubation time of 18 hours. A convolutional neural network (CNN) trained on optical images of the platform was able to classify samples as contaminated or uncontaminated with a validation accuracy of over 93%.