Engineered Spores as Fluorogenic Biological Indicators for Sterility Testing

描述（由申请人提供）：BCR Diagnostics，Inc. (BCR)最近开发了一种技术平台并获得专利，用于将荧光功能工程化到活芽孢杆菌孢子中。在BCR产生的荧光孢子（F-孢子）在生理上似乎与正常孢子相同，除了作为初始萌发事件的自我报告者。考虑到F-孢子的独特特征以及目前对无菌试验快速可靠生物指示剂（BI）的需求，BCR提出了蒸汽灭菌快速生物指示剂（rBI）的II期确认研究。本研究的总体目标是建立一个数据库，以便向美国食品药品监督管理局提交上市前510（k）通知。 该应用的关键是灭菌过程对医疗保健行业至关重要，并且需要监测所用设备和程序的有效性。目前，活微生物孢子的生物指示剂是最准确和可靠的无菌系统，但这些指示剂的主要缺点是，表面上无菌的物品必须储存很长时间才能确保无菌。 拟议的生物指示剂应与常规指示剂一样发挥作用，因为两者都依赖于生命功能作为细胞死亡的参数。然而，BCR的指示剂是不同的，因为它测量早期萌发事件，而常规指示剂测量孢子生长到细菌培养物中。 本研究包含三个主要任务：任务1。设计并构建二维侧向流系统原型，以测量暴露于灭菌周期后rBI的荧光活性。对于任务1，BCR将与New Horizons Diagnostics Corp.（哥伦比亚，MD）合作。任务2.标准化并测试RBI的稳定性和重现性。任务3.进行：双盲：研究期间，将通过将rBI与传统BI进行比较（使用孢子生长作为参数）来评价rBI的性能（有效性）。对于任务3，BCR将与Raven Biological Laboratories（Omaha，NE）合作。 公共卫生相关性：灭菌在许多行业中至关重要，从医疗保健和制药到发酵和食品服务。然而，测试灭菌过程有效性的常规技术涉及基于培养的监测，其可能需要长达七天才能完成。为了提供更快速的无菌保证，BCR Diagnostics设计了独特的活微生物，当灭菌过程不充分时，这些微生物会迅速（不到30分钟）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 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.