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Proceedings Paper

Development of bacteriophage-based bioluminescent bioreporters for monitoring of microbial pathogens
Author(s): Aysu Ozen; Kacey Montgomery; Pat Jegier; Stacey Patterson; Kathleen A. Daumer; Steven A. Ripp; Jay L. Garland; Gary S. Sayler
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Paper Abstract

Microorganisms pose numerous problems when present in human occupied enclosed environments. Primary among these are health related hazards, manifested as infectious diseases related to contaminated drinking water, food, or air circulation systems or non-infectious allergy related complications associated with microbial metabolites (sick building syndrome). As a means towards rapid detection of microbial pathogens, we are attempting to harness the specificity of bacterial phage for their host with a modified quorum sensing amplification signal to produce quantifiable bioluminescent (lux) detection on a silicon microluminometer. The bacteriophage itself is metabolically inactive, only achieving replicative capabilities upon infection of its specific host bacterium. Bacteriophage bioluminescent bioreporters contain a genomically inserted luxI component. During an infection event, the phage genes and accompanying luxI construct are taken up by the host bacterium and transcribed, resulting in luxI expression and subsequent activation of a homoserine lactone inducible bioluminescent bioreporter. We constructed a vector carrying the luxI gene under the control of a strong E. coli promoter and cloned it into E. coli. We have shown that it can induce luminescence up to 14,000 counts per second when combined with the bioreporter strain. In their final embodiment, these sensors will be fully independent microelectronic monitors for microbial contamination, requiring only exposure of the biochip to the sample, with on-chip signal processing downloaded directly to the local area network of the environmental control system.

Paper Details

Date Published: 4 March 2004
PDF: 11 pages
Proc. SPIE 5270, Environmental Monitoring and Remediation III, (4 March 2004); doi: 10.1117/12.516181
Show Author Affiliations
Aysu Ozen, Univ. of Tennessee/Knoxville (United States)
Kacey Montgomery, Univ. of Tennessee/Knoxville (United States)
Pat Jegier, Univ. of Tennessee/Knoxville (United States)
Stacey Patterson, Univ. of Tennessee/Knoxville (United States)
Kathleen A. Daumer, Dynamac Corp. (United States)
Steven A. Ripp, Univ. of Tennessee/Knoxville (United States)
Jay L. Garland, Dynamac Corp. (United States)
Gary S. Sayler, Univ. of Tennessee/Knoxville (United States)

Published in SPIE Proceedings Vol. 5270:
Environmental Monitoring and Remediation III
Tuan Vo-Dinh; Klaus P. Schäfer; Guenter Gauglitz; Dennis K. Killinger; Robert A. Lieberman, Editor(s)

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