HOMOGENEOUS ASSAY FOR SPECIFIC DNA/RNA SEQUENCE BY FRET

APPLICANT?S DESCRIPTION: The proposed research project will develop a homogeneous assay platform for nucleic acid diagnostics that brings improvements based on ease of use (a true "mix and read" format) and quantification, while approaching the sensitivity possible with the much more cumbersome PCR assay. The immediate focus of this research will be development of assays for pathogenic bacterial infection, using E.coli serotype 017:H7 as the initial target. The format will be applied in Phase II to other pathogenic bacteria and to oncogenic viruses. The use of large complexes as the fluorescent donors and acceptors will provide sensitivity not achieved with present homogeneous assays and enable the use of this format in miniaturized assays and high-density arrays. PROPOSED COMMERCIAL APPLICATION: Food-borne pathogens are of major concern in the United States and worldwide. The food-borne pathogen detection market is predicted to reach $178 million dollars by the year 2005. This manifests a need for a quick, inexpensive, sensitive detection system that can identify the pathogenic bacteria regardless of the source.