Development of Detection Technologies for Pathodgens and Toxins and their Validation in Food Matrices

PPROACH: Development of adequate sampling protocols is a critical component of any detection scheme. Methods will be developed that result in separation of pathogens from an aggregate matrix or biofilm, and simultaneously enrich and concentrate multiple pathogenic species. Extraction and concentration is essential for a biosensor that does not rely on a preenrichment culture step. Pathogens will be prepared and concentrated using techniques such as sequential filtration, gravitation, immunotrapping, ligand-receptor interactions, and cell sorting. If preenrichment is necessary, methods will be developed to shorten the incubation times required. Methods such as integrated optical sensing, mass spectrometry, sensitive fluorescent, immuno-electrochemical detection, immunomagnetic electrochemiluminescence, microchip arrays, and combinatorial chemistry will be adapted to meet these goals. Reagents, such as monoclonal antibodies, receptors, and molecular imprints will be generated to facilitate separation and detection schemes. FY01 Program Increase $269,370. 1SY Added. Bridging project replacing 5325-42000-027-00D (Feb. 05) FY05 Prog. Inc. %400,000. Add 1 SY.
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PROGRESS: 2005/01 TO 2005/12
Progress Report 1. What major problem or issue is being resolved and how are you resolving it (summarize project aims and objectives)? How serious is the problem? Why does it matter? This project addresses the need for validated methods to detect toxins and pathogens in foods. Existing surveillance systems are designed to detect occurrences of infections and intoxications resulting from consumption of foods naturally contaminated with pathogens or the toxins produced by these organisms. However, it has been of increasing concern that the intentional adulteration of food with chemical or biological agents such as bacterial and plant toxins could become a major instrument of bioterrorism. In addition, it is likely that crude, rather than purified, toxins would be used as bioweapons, and it is possible that combinations of toxins could be employed. Therefore, some data gaps related to dose response relationships must be filled. In addition, rapid, portable sample preparation techniques applicable to important food matrices are needed, particularly for new detection technologies. This research is administered under National Program 108 Food Safety. <P>
2. List by year the currently approved milestones (indicators of research progress) Year 1 (FY03) Obtain Campylobacter and Mycobacteria strains and immunize mice Year 2 (FY04) New ELISA for Campylobacter strain analysis New ELISA for Mycobacteria strain analysis Year 3 (FY05) Validation of new Campylobacter and Mycobacteria strain-specific ELISAs Year 4 (FY06) [None] This project was terminated and replaced by 5325-32000-043-00D.<P> 4a List the single most significant research accomplishment during FY 2006. This project has been replaced by 5325-32000-043-00D. 4d Progress report. This bridging CRIS project, replacing CRIS 5325-42000-027-00D in FY05, terminated in FY06. It was replaced with Project 5325-42000-043-00D Development of Detection Technologies for Toxins and their Validation in Food Matrices, which has a separate report. Milestones listed for question 2 and 3A refer to old Project 5325-42000-027-00D. Milestones listed for question 3B are from the new Project 5325-42000-043-00D, which underwent OSQR in FY06. The Major Accomplishment described for question 5 and Technology Transfer in question 6 took place under Project 5325-42000- 027-00D. <P>5. Describe the major accomplishments to date and their predicted or actual impact. Upon a request from the Food Safety Inspection Service (FSIS) of the USDA, we developed an assay for residues of Ceftiofur, a broad spectrum cephalosporin antibiotic used in beef production. The immunoassay we created is faster and cheaper than previous bioassay methods and is now routinely employed by FSIS for regulation. (This work was performed under CRIS 5325-42000-027-00D.)<P>
6. What science and/or technologies have been transferred and to whom? When is the science and/or technology likely to become available to the end- user (industry, farmer, other scientists)? What are the constraints, if known, to the adoption and durability of the technology products? We have developed an assay for residues of Ceftiofur, a regulated antibiotic found in beef. Our immunoassay is faster and cheaper than previous methods. In FY04 technology transfer to FSIS was formally completed, along with reagents and training/consultation. (This work was performed under CRIS 5325-42000-027-00D.) Immunoassay technology for Bowman-Birk Inhibitor, a naturally-occurring anti-nutritional component of soybeans, was originally developed under a separate project. Transferred to this Project, the technology has now been patented and licensed to an immunodiagnostics manufacturer, Agdia, Inc. (Elkhart, IN). (This work was performed under CRIS 5325-42000-027- 00D.)