Rapid, Low-Cost System for Detection of Bacterial Contamination of Blood Pla--OLD

描述（由申请人提供）：这个STTR第一期项目将证明一种低成本、快速检测血小板浓缩物中细菌污染的系统的概念，只需几分钟，而不是几天，成本/性能等于公认的测试方法。尽管美国是世界上血液供应最安全的国家之一，但输入被致病菌污染的血液制品仍存在重大危险。在美国，除人为失误外，细菌感染是最危险的输血相关风险。血库培养血小板2天进行细菌检测。由于血小板单位的保质期为5天，这2天的保质期减少对供应有严重的影响。该解决方案将使血小板寿命延长30%以上，并且能够释放比旧血小板更安全、更有效的新鲜血小板。这项研发工作的目标是建立和测试一个概念验证系统，并开发一种用于检测血小板浓缩物中细菌的染色方法。提出的专有方法被称为喷泉流式细胞术。这种方法是基于一种系统，其中含有微生物的溶液流，用荧光探针标记，用LED照亮，用数码相机检测荧光。使用低成本的商用数码相机和led使得这种设备在经济上具有吸引力。这项拟议中的研究将用一种系统测试不同的染料组合，该系统被设计用来检测在红细胞和白细胞背景下插入单采血小板的细菌。预期的结果是替代更昂贵、灵敏度更低、速度更慢的检测系统。喷泉流”将允许检测血小板污染在自动培养装置的灵敏度，但在~10分钟。该系统可以自动化，并且可能允许每天检查数百个血小板单位。试验将在大肠杆菌接种到人单采血小板单位上进行。关键绩效标准将是大肠杆菌的方法检测限（MDL）、总测定时间、检测效率和假阳性检测率。为了证明该技术的商业可行性，我们的总体性能目标是MDL为~1个细菌/ml（在10ml样品中），检测效率为>75%（检测到的细菌/存在的细菌），流速为>3.0 ml/分钟。该项目的关键任务是开发一种能够实现这些目标的染色试剂组合。这项研究将由SoftRay公司的一组科学家与其怀俄明大学STTR合作伙伴、输血医学顾问（丹佛邦菲尔斯血液中心）和临床微生物学家（克利夫兰诊所）合作进行。公共卫生相关性：在美国，每年大约输注400万单位的血小板浓缩物，估计有千分之一到三千分之一的单位被潜在致病菌污染。在美国，被细菌污染的血液是除人为失误外最危险的输血风险。本项目提出的系统将提供更低成本、更及时和更敏感的细菌污染检测，为输血提供更安全的血小板供应。

DESCRIPTION (provided by applicant): This STTR Phase I project will prove the concept for a low-cost, rapid system for detecting bacterial contamination in platelet concentrates, in minutes and not days, with cost/performance equal to accepted testing methodologies. Though the US has one of the safest blood supplies in the world, significant danger exists from the transfusion of blood products contaminated with pathogenic bacteria. Bacterial infection represents the most dangerous transfusion-related risk in the US other than human error. Blood banks incubate platelet cultures for 2 days for bacterial testing. Since platelet units have a shelf life of 5 days, this 2- day reduction in shelf life has a harsh impact on the supply. The proposed solution will extend platelet lifetime by more than 30%, as well as enable the release of fresher platelets, which are safer and more effective than older platelets. The objective of this proposed R&D effort is to build and test a proof-of-concept system and develop a staining method for the detection of bacteria in platelet concentrate units. The proposed proprietary method is called Fountain Flow" Cytometry. This method is based on a system in which a stream of solution containing the microorganisms, labeled with fluorescent probes, is illuminated with an LED and the fluorescence is detected with a digital camera. The use of low-cost commercial digital cameras and LEDs makes this device economically attractive when compared to alternatives. The proposed research will test alternative dye combinations with a system designed to detect bacteria spiked into apheresis platelets against a background of red and white blood cells. The anticipated result is an alternative to more expensive, less sensitive, and slower detection systems. Fountain Flow" will allow detection of platelet contamination at the sensitivity of automated culturing devices, but in ~10 minutes. The system can be automated and would potentially allow the examination of hundreds of platelet units per day. Tests will be performed on E. coli inoculated into human apheresis platelet units. Key performance criteria will be the method detection limit (MDL) for E. coli, total assay time, detection efficiency, and rate of false- positive detections. Our overall performance objectives to demonstrate commercial viability of the technique are an MDL of ~1 bacterium/ml (in a 10ml sample) with a detection efficiency >75% (bacteria detected/bacteria present) and a flow rate of >3.0 ml/minute. The key task in this project is to develop a combination of staining reagents that will achieve these objectives. This research will be performed by a team of scientists from SoftRay Inc. in colaboration with its University of Wyoming STTR partner, a transfusion medicine consultant (Bonfils Blood Center, Denver), and a clinical microbiologist (Cleveland Clinic). PUBLIC HEALTH RELEVANCE: Approximately four million units of platelet concentrates are transfused in the United States annually, with an estimated one in one thousand to one in three thousand units contaminated with potentially pathogenic bacteria. Blood contaminated by bacteria now represents the most dangerous transfusion related risk in the US other than human error. This project proposes a system that will provide for lower-cost, more timely and more sensitive detection of bacterial contamination, providing for a safer blood platelet supply for transfusion.