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This proposal describes a molecular fluorescent-based assay that simultaneously detects up to five possible enterotoxin genes that may be found in Bacillus strains in contaminated food. The assay uses gene-specific fluorescent tags that allow for quantitative real-time detection of enterotoxin genes in contaminated food in a fraction of the time required for alternative DNA-based tests.
The development of molecular-based methods to ensure food safety has significantly improved the speed and sensitivity with which foodborne pathogens may be detected/enumerated. One food pathogen of recent concern is the spore-forming bacterium Bacillus cereus. The proposed project targets this pathogen and other closely related species that harbor enterotoxin genes. If present at high levels in certain foods, such as dairy products, sauces, or rice, enterotoxigenic Bacillus spp. elicit diarrheal symptoms that persist for several hours. The presence of viable cells/spores may not noticeably alter the flavor or appearance of affected foods. In some cases, such as contaminated infant milk formula, illness may be quite severe. This proposal describes a molecular fluorescent-based assay that simultaneously detects up to five possible enterotoxin genes that may be found in Bacillus strains in contaminated food. The assay uses gene-specific fluorescent tags that allow for quantitative real-time detection of enterotoxin genes in contaminated food in a fraction of the time required for alternative DNA-based tests. Detection and confirmation of target pathogens are completed simultaneously, reducing costs associated with multiple steps. The application of this system for pathogen detection in foods will expand the scope of tests currently available, and will positively impact agriculture by increasing the rate of quality control steps for foods during processing.