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Proceedings Paper

A model system for pathogen detection using a two-component bacteriophage/bioluminescent signal amplification assay
Author(s): Nathan G. Bright; Richard J. Carroll; Bruce M. Applegate
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Paper Abstract

Microbial contamination has become a mounting concern the last decade due to an increased emphasis of minimally processed food products specifically produce, and the recognition of foodborne pathogens such as Campylobacter jejuni, Escherichia coli O157:H7, and Listeria monocytogenes. This research investigates a detection approach utilizing bacteriophage pathogen specificity coupled with a bacterial bioluminescent bioreporter utilizing the quorum sensing molecule from Vibrio fischeri, N-(3-oxohexanoyl)-homoserine lactone (3-oxo-C6-HSL). The 3-oxo-C6-HSL molecules diffuse out of the target cell after infection and induce bioluminescence from a population of 3-oxo-C6-HSL bioreporters (ROLux). E. coli phage M13, a well-characterized bacteriophage, offers a model system testing the use of bacteriophage for pathogen detection through cell-to-cell communication via a LuxR/3-oxo-C6-HSL system. Simulated temperate phage assays tested functionality of the ROLux reporter and production of 3-oxo-C6-HSL by various test strains. These assays showed detection limits of 102cfu after 24 hours in a varietry of detection formats. Assays incorporating the bacteriophage M13-luxI with the ROLux reporter and a known population of target cells were subsequently developed and have shown consistent detection limits of 105cfu target organisms. Measurable light response from high concentrations of target cells was almost immediate, suggesting an enrichment step to further improve detection limits and reduce assay time.

Paper Details

Date Published: 30 March 2004
PDF: 7 pages
Proc. SPIE 5271, Monitoring Food Safety, Agriculture, and Plant Health, (30 March 2004); doi: 10.1117/12.547803
Show Author Affiliations
Nathan G. Bright, Purdue Univ. (United States)
Richard J. Carroll, Purdue Univ. (United States)
Bruce M. Applegate, Purdue Univ. (United States)

Published in SPIE Proceedings Vol. 5271:
Monitoring Food Safety, Agriculture, and Plant Health
George E. Meyer; Yud-Ren Chen; Shu-I Tu; Bent S. Bennedsen; Andre G. Senecal, Editor(s)

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