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Article

An Engineered Reporter Phage for the Fluorometric Detection of Escherichia coli in Ground Beef

1
Department of Food Science, Cornell University, Ithaca, NY 14853, USA
2
Department of Food Science and Technology, The Ohio State University, Columbus, OH 43210, USA
3
Department of Biological Systems Engineering, Virginia Tech, Blacksburg, VA 24061, USA
*
Author to whom correspondence should be addressed.
Academic Editor: Maria Serena Chiucchi
Microorganisms 2021, 9(2), 436; https://doi.org/10.3390/microorganisms9020436
Received: 2 February 2021 / Revised: 11 February 2021 / Accepted: 17 February 2021 / Published: 19 February 2021
Despite enhanced sanitation implementations, foodborne bacterial pathogens still remain a major threat to public health and generate high costs for the food industry. Reporter bacteriophage (phage) systems have been regarded as a powerful technology for diagnostic assays for their extraordinary specificity to target cells and cost-effectiveness. Our study introduced an enzyme-based fluorescent assay for detecting the presence of E. coli using the T7 phage engineered with the lacZ operon which encodes beta-galactosidase (β-gal). Both endogenous and overexpressed β-gal expression was monitored using a fluorescent-based method with 4-methylumbelliferyl β-d-galactopyranoside (MUG) as the substrate. The infection of E. coli with engineered phages resulted in a detection limit of 10 CFU/mL in ground beef juice after 7 h of incubation. In this study, we demonstrated that the overexpression of β-gal coupled with a fluorogenic substrate can provide a straightforward and sensitive approach to detect the potential biological contamination in food samples. The results also suggested that this system can be applied to detect E. coli strains isolated from environmental samples, indicating a broader range of bacterial detection. View Full-Text
Keywords: bacteria detection; engineered bacteriophages; fluorescent assay; beta-galactosidase; ground beef bacteria detection; engineered bacteriophages; fluorescent assay; beta-galactosidase; ground beef
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MDPI and ACS Style

Chen, A.; Wang, D.; Nugen, S.R.; Chen, J. An Engineered Reporter Phage for the Fluorometric Detection of Escherichia coli in Ground Beef. Microorganisms 2021, 9, 436. https://doi.org/10.3390/microorganisms9020436

AMA Style

Chen A, Wang D, Nugen SR, Chen J. An Engineered Reporter Phage for the Fluorometric Detection of Escherichia coli in Ground Beef. Microorganisms. 2021; 9(2):436. https://doi.org/10.3390/microorganisms9020436

Chicago/Turabian Style

Chen, Anqi, Danhui Wang, Sam R. Nugen, and Juhong Chen. 2021. "An Engineered Reporter Phage for the Fluorometric Detection of Escherichia coli in Ground Beef" Microorganisms 9, no. 2: 436. https://doi.org/10.3390/microorganisms9020436

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