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, pre-detection culture enrichment.
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 simple stability for light addressable potentiometric detection;
<li>A cytolethaldistending toxin (CDT)-mediated membrane disintegration assay;
<li>A liquid crystal detection system. </ol>
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.