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United States Department of Agriculture

Agricultural Research Service

Research Project: ASSAY DEVELOPMENT FOR DETECTION OF PATHOGENIC E. COLI IN ENVIRONMENTAL SAMPLES

Location: Environmental Microbial and Food Safety Laboratory

2012 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, a spectrofluoremeter instrument (Signalyte) was developed based on the Integrating Waveguide Biosensor. Under the current agreement, an immunoassay was developed for the rapid and sensitive detection of pathogenic E. coli. Briefly, cells were captured using immunomagnetic beads and then tagged with a fluorescent-labeled antibody. Cells were then quantified using the Signalyte. As few as ten cells were detected per ml of water. Rapid, sensitive methods for detection of water-borne pathogens are critical for understanding fate and transport processes.


Last Modified: 8/21/2014
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