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Bioluminescent Imaging for High Throughput Screening for Bacterial Pathogens and Toxins

Award No.: 
N-00014-04-1-0659
Status: 
Complete
Principal Investigator: 
Mansel Griffiths
PI Organization: 
University of Guelph
Abstract: 
The goal of the project is to develop cell-based bioluminescent biosensor for pathogenicity. The following objectives were planned: 1. Amass a collection of bioluminescent bacteria, both pathogenic and non-pathogenic, and investigate their attachment to mammalian cell monolayers, and to subsequently correlate attachment with pathogenicity. 2. Construct mammalian cell cultures carrying firefly luciferase and aequorin or obelin genes, establish correlation between the level of bioluminescence and intracellular concentrations of Ca 2+ and ATP. 3. Investigate the changes in both Ca 2+ and ATP in mammalian cells after interaction with pathogenic and non-pathogenic bacteria, establish the cut-off level of bioluminescence that separates pathogens from non-pathogens.
Analytic Methods Utilized: