2009 Annual Report 1a.Objectives (from AD-416) The objective of this cooperative research project is to develop an assay for the detection of pathogenic E. coli in water, soil, manure and food matrices using an integrated immuno-PCR biosensor. 1b.Approach (from AD-416) Pathogenic E. coli will be captured and fluorescently labeled with specific monoclonal antibodies. The E. coli cell-antibody complex will subsequently be detected using an immunoassay biosensor followed by PCR characterization. 3.Progress Report Food-borne and water-borne pathogens pose a serious threat to public health. Under a previous CRADA, instrumentation (Signalyte TM) was developed based on the Integrating Waveguide Biosensor. Under the current agreement, an immunoassay is being developed for the rapid and sensitive detection of pathogenic E. coli. Briefly, cell are incubated with fluorescent-labeled antibodies, the cells filtered to remove excess un-reacted antibody, the antibodies released with a brief acidic treatment, and the antibody concentration subsequently quantified utilizing a spectrofluoremeter (Signalyte TM). As few as 100 cells can be detected in 30-60 minutes. Meetings are held bi-monthly to discuss results with frequent email communication in the interim.