Viability Assay for Monitoring Decontamination of Pathogenic Bacteria

Not Available "To determine the effectiveness of antimicrobial agents in decontamination and antimicrobial efficacy testing, the plate count method has been traditionally used. This method requires trained technical expertise, large amounts of microbiological growthmedia and up to several days to complete and often with unreliable results. Luna Innovations proposes to develop a rapid, automated means of determining the efficacy of antimicrobial agents on vegetative bacterial cells using fluorescent probes andantibiotics targeting the primary susceptibility regions of tester organisms (e.g., cell wall, cell membrane and cellular respiration). In the Phase I work, the fluorescence-based formats will focus on measuring changes in pH, redox and cell metabolism toquantify the amount of viable organisms remaining after treatment with the decontaminants and determine the kinetics associated with the cell destruction. Fluorescence-based responses obtained in 5-10 minutes will be compared to traditional plate countdeterminations as controls. Tester strains will include ampicillin+/- and b-galactosidase+/- strains of Escherichia coli and protein A+/- strains of Staphyloccus aereus. In subsequent Phase II studies, the types of tester organisms will be expanded toinclude other pathogenic microorganisms and indicator compounds. In addition, the instrumentation will be adapted to have multiple sampling and the high throughput capabilities. The introduction of the concept of viable but nonculturable cells in the1980s has led to important research work concerning the existence and significance of these kinds of cells within bacterial communities. In addition, quantification of total bacterial numbers in a fast and reliable fashion is a basic and essential task inseveral areas of microbiology, including public health, biotechnology, food and water industries, pharmaceuticals and the natural environment. Our proposed technology would greatly increase the speed and reliability with which organisms could be assessedafter a decontamination event both in the labor