The need to identify multiple targets for measurement and monitoring is common to many problems in the detection of human, fish, and shellfish pathogens. We propose to integrate knowledge and instrumentation for the purpose of: (a) allowing periodic broad-scope assessment of entire panels of pathogens; (b) increasing both flexibility and specificity in the targeting of organisms for routine testing; and (c) diminishing the cost of surveillance. Products of DNA amplification of a given genetic locus from all members of an assemblage will be examined with increasingly automated systems that can separate such sequences and permit rapid generation of sequence information diagnostic of individual members of evolutionary-related organisms. The powerful Denaturing High Performance Liquid Chromatography platform (Transgenomic technology) will be tested for the separation of highly similar PCR products, and automated peak recovery will allow rapid acquisition of sequence information. COMMERCIAL APPLICATIONS: Potential commercial applications exist in fish and shellfish health monitoring, environmental conditions surveillance in culture systems, and in seafood safety assurance. At the successful conclusion of Phase 1 research, we expect to have demonstrated that whole groups of microorganism (microbial consortia) can be characterized by techniques either partially automated at this time or subject to automation in Phase 2. We anticipate that by the end of Phase 2, hybridization probes designed from sequences obtained either by DGGE or D-HPLC fractions will be able to be used to conduct assays for microbial pathogens in the field, using portable instruments now under development by our research partners.
