Engineered Spores as Fluorogenic Biological Indicators for Sterility Testing

描述(由申请人提供)：BCR诊断公司(BCR)最近开发并申请了一项技术平台，用于将荧光功能工程到活的芽孢杆菌孢子中。在BCR产生的荧光孢子(F孢子)似乎在生理上与正常孢子相同，只是作为初始萌发事件的自我报告者。 考虑到F孢子的独特特性以及现有对用于无菌检测的快速可靠的生物指示器(BIS)的需求，BCR建议进行第一阶段研究，以评估使用F孢子开发用于无菌保证的快速生物指示器的可行性。 对应用至关重要的是，灭菌过程对医疗保健行业至关重要，并需要监测所用设备和程序的有效性。目前，活的微生物孢子的生物指示器是无菌物品最准确和最可靠的系统，但这些指示器的主要缺点是，似乎已灭菌的物品必须长期储存，以确保无菌。建议的生物指标应与传统指标一样发挥作用，因为两者都依赖于生命功能作为细胞死亡的参数。然而，BCR的指标是不同的，因为它衡量的是早期萌发事件，而传统指标衡量的是孢子长入细菌培养物中。拟议的研究包括三个主要任务：任务1。优化F孢子作为蒸汽灭菌后无菌保证的生物学指标。生物指示器原型将被生产、标准化，然后进行稳定性和重复性测试。任务2：以孢子生长为参数，将生物指标原型与常规生物指标进行比较，对生物指标原型进行评价。任务3.进行旨在扩大F孢子在使用环氧乙烷、过氧化氢气体等离子体和辐射灭菌过程中的使用的实验。 从医疗保健和制药到发酵和食品服务，灭菌在许多行业都是至关重要的。然而，测试灭菌过程有效性的传统技术涉及基于培养的监测，可能需要长达四天的时间才能完成。为了提供更快速的无菌保证，BCR Diagnostics开发了独特的活微生物，当灭菌过程不充分时，这种微生物会迅速发出荧光信号。

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.