Engineered Spores as Fluorogenic Biological Indicators for Sterility Testing

DESCRIPTION (provided by applicant): 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 I study to evaluate the feasibility of using F-spores to develop a rapid biological indicator for sterility assurance. 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 articles 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. Optimize F- spores as biological indicators for sterility assurance after steam sterilization. A biological indicator prototype will be produced, standardized and then tested for stability and reproducibility. Task 2. Evaluate the biological indicator prototype by comparing it with conventional biological indicators using spore outgrowth as a parameter. Task 3. Conduct experiments designed to extend the use of F-spores for sterilization processes using ethylene oxide, hydrogen peroxide gas plasma, and radiation. 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 four days to complete. To provide more rapid sterility assurance, BCR Diagnostics has developed unique living microorganisms that promptly send fluorescent light signals when the sterilization process is inadequate.