BCR Diagnostics, Inc. (BCR) has recently developed and patented a technology platform for engineering fluorogenic functionality into living Bacillus spores. Fluorogenic spores (F-spores) produced at BCR appear to be physiologically identical to normal spores except for performing as self reporters of initial germination events. Considering the unique features of the F- spores in context with existing needs for rapid and reliable biological indicators (BIs) for sterility testing, BCR proposes a Phase II validation study of a rapid biological indicator (rBI) for steam sterilization. The study's overall objective is to assemble a database for submitting a premarket 510(k) notification to the Food and Drug Administration. Critical to the application is that sterilization processes are essential to the healthcare industry and are required to monitor the effectiveness of equipment and procedures used. Currently, biological indicators of living microbial spores are the most accurate and reliable systems for sterility but the major disadvantage with these indicators is that seemingly sterilized articles must be stored for prolonged times to ensure sterility. The proposed biological indicator should perform as the conventional indicators because both depend on vital functions as parameters of cell death. However, BCR's indicator is different because it measures early germination events while conventional indicators measure spore outgrowth into a bacterial culture. The proposed study consists of three major assignments: Task 1. To design and construct a prototype of a 2-dimensional lateral flow system to measure fluorogenic activity of the rBI after exposure to a sterilization cycle. For Task 1, BCR will collaborate with New Horizons Diagnostics Corp. (Columbia, MD). Task 2. To standardize and test the rBI for stability and reproducibility. Task 3. To conduct a :double blind: study during which the performance (efficacy) of the rBI will be evaluated by comparing it to a conventional BI using spore outgrowth as parameter. For Task 3, BCR will collaborate with Raven Biological Laboratories (Omaha, NE). PUBLIC HEALTH RELEVANCE: Sterilization is critical in a number of industries, from healthcare and pharmaceuticals to fermentation and food service. Conventional techniques to test the effectiveness of sterilization processes, however, involve culture based monitoring that can take up to seven days to complete. To provide more rapid sterility assurance, BCR Diagnostics has engineered unique living microorganisms that promptly (less than 30 min) send fluorescent light signals when the sterilization process is inadequate.