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孢子在现有无菌检测中需要快速可靠的生物指示器(BIS)的独特特性，BCR建议对蒸汽灭菌的快速生物指示器(RBI)进行第二阶段验证研究。这项研究的总体目标是建立一个数据库，用于向食品和药物管理局提交上市前510(K)通知。 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最准确和可靠的无菌系统，但这些指示器的主要缺点是，看似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。 这项拟议的研究包括三个主要任务：任务1.设计和建造一个二维侧向流动系统的原型，以测量暴露在灭菌周期中的RBI的荧光活性。对于任务1，BCR将与新视野诊断公司(马里兰州哥伦比亚市)合作。任务2.标准化和测试RBI的稳定性和重复性。任务3.进行：双盲：研究，在此期间，将以孢子生长为参数，通过将其与传统的BI进行比较来评价RBI的性能(效果)。对于任务3，BCR将与乌鸦生物实验室(内华达州奥马哈)合作。 与公共健康相关：从医疗保健和制药到发酵和食品服务，绝育在许多行业中都是至关重要的。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7天的时间才能完成。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，这种微生物会迅速(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.