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The specific objectives of this research are to develop: <ol> <li>Rapid and effective methods to separate and concentrate various pathogens from food samples in order to shorten the duration of, or reduce the need for, culture enrichment;
<li>Simple and effective sensors for the screening detection of food-borne pathogens;
<li>Integrated detection procedures suitable for regulatory and/or industrial applications.</ol>
To develop the means to select and concentrate only target organisms in the presence of an elaborate non-pathogenic microflora. We propose to develop both IMB and immuno-affinity column technologies to more rapidly concentrate various food-borne target pathogens and thereby shorten the time period of, and minimize the need for, predetection culture enrichment.
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Biosensor methods will also be developed including, but not limited to: <ol>
<li>An alkaline phosphatase-antibody system suitable for 96-well microplate readers;
<li>Metabolic control of cellular ATP level to confirm the presence of viable cells;
<li>The use of the streptavidin-biotin interaction to enhance sample stability for light addressable potentiometric detection;
<li>A cytolethal distending toxin (CDT) - mediated membrane disintegration assay; and
<li>A liquid crystal detection system.</ol>
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Lastly, we propose to integrate and combine the most promising of these procedures into one, or more, screening protocols for regulatory and/or industrial applications.