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Biomolecules 2013, 3(3), 563-577; doi:10.3390/biom3030563

A Sensitive DNA Enzyme-Based Fluorescent Assay for Bacterial Detection

1
Department of Biochemistry and Biomedical Sciences, McMaster University, 1280 Main St. W., Hamilton, ON L8S 4K1, Canada
2
Dvision of Gastroenterology, Department of Medicine, McMaster University, 1280 Main St. W., Hamilton, ON L8S 4K1, Canada
3
Department of Chemistry and Chemical Biology, McMaster University, 1280 Main St. W., Hamilton, ON L8S 4K1, Canada
4
Michael G. DeGroote Institute for Infectious Disease Research, McMaster University, 1280 Main St. W., Hamilton, ON L8S 4K1, Canada
These authors contributed equally to this work.
*
Author to whom correspondence should be addressed.
Received: 25 July 2013 / Revised: 9 August 2013 / Accepted: 11 August 2013 / Published: 20 August 2013
(This article belongs to the Special Issue Enzymes and Their Biotechnological Applications)
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Abstract

Bacterial detection plays an important role in protecting public health and safety, and thus, substantial research efforts have been directed at developing bacterial sensing methods that are sensitive, specific, inexpensive, and easy to use. We have recently reported a novel “mix-and-read” assay where a fluorogenic DNAzyme probe was used to detect model bacterium E. coli. In this work, we carried out a series of optimization experiments in order to improve the performance of this assay. The optimized assay can achieve a detection limit of 1000 colony-forming units (CFU) without a culturing step and is able to detect 1 CFU following as short as 4 h of bacterial culturing in a growth medium. Overall, our effort has led to the development of a highly sensitive and easy-to-use fluorescent bacterial detection assay that employs a catalytic DNA.
Keywords: bacterial detection; DNAzyme; fluorescence; biosensor; Escherichia coli bacterial detection; DNAzyme; fluorescence; biosensor; Escherichia coli
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This is an open access article distributed under the Creative Commons Attribution License (CC BY 3.0).

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MDPI and ACS Style

Aguirre, S.D.; Ali, M.M.; Salena, B.J.; Li, Y. A Sensitive DNA Enzyme-Based Fluorescent Assay for Bacterial Detection. Biomolecules 2013, 3, 563-577.

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