Ms. Aysu Ozen Profile

Aysu Ozen

at Univ of Tennessee

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Proceedings Article | 14 June 2004 Paper

Phage-amplified bioluminescent bioreporters for the detection of foodborne pathogens

Steven Ripp, Jacque Young, Aysu Ozen, Patricia Jegier, Courtney Johnson, Kathleen Daumer, Jay Garland, Gary Sayler

Proceedings Volume 5329, (2004) https://doi.org/10.1117/12.529436

KEYWORDS: Pathogens, Bioluminescence, Biosensing, Integrated circuits, Signal processing, Biological detection systems, Environmental sensing, Organisms, Signal generators, Industrial chemicals

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The objective of this investigation is to develop a bioluminescent bioreporter system for the detection and monitoring of pathogenic microbial species. Current detection methodologies typically rely on time-consuming sample pre-enrichment steps to elevate pathogen concentrations to detectable levels or DNA based polymerase chain reaction (PCR) techniques that require extensive user training and expensive instrumentation. Detection utilizing bioluminescent bioreporter organisms, however, can provide a simple and rapid means of monitoring foodborne pathogens. Bioluminescent bioreporters are engineered to produce light in response to specific environmental inducers. The light signal is then measured with photodetector devices to generate a quantitative assessment of inducer concentration. The immediate goal of this research effort is to integrate key quorum sensing signal transduction elements into pathogen specific bacteriophages. Upon infection of a unique pathogenic species by the bacteriophages, quorum sensing signals will be generated that will subsequently stimulate bioluminescence in neighboring bioluminescent bioreporter cells. Utilizing both bacteriophages and bioluminescent bioreporters, we realize exceptional pathogen specificity while attaining enhanced bioluminescence production. This integrative approach will lead to rapid pathogen identification without requisite sample pre-enrichment. Additionally, since the bioluminescent response is completely intrinsic to the bioreporter organism, no user interventions are required for generating light signals; the protocol requires only addition of the food sample with the bacteriophage/bioluminescent bioreporter system. Measurement of light responses can be achieved using high-throughput microtiter plate readers, hand-held photomultiplier units, or microchip luminometers.

Proceedings Article | 4 March 2004 Paper

Development of bacteriophage-based bioluminescent bioreporters for monitoring of microbial pathogens

Aysu Ozen, Kacey Montgomery, Pat Jegier, Stacey Patterson, Kathleen Daumer, Steven Ripp, Jay Garland, Gary Sayler

Proceedings Volume 5270, (2004) https://doi.org/10.1117/12.516181

KEYWORDS: Pathogens, Soil contamination, Signal processing, Contamination, Environmental monitoring, Water, Organisms, Luminescence, Environmental sensing, Bioluminescence

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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.

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