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

快速、低成本检测血液中细菌污染的系统--OLD

• 批准号: 7748667
• 负责人: Scott Irwin
• 金额: $ 10.96万
• 依托单位: SOFTRAY, INC.
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-16 至 2010-09-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7748667
• 关键词: Address; Bacteria; Bacterial Infections; Biological Assay; Blood; Blood Banks; Blood Component Removal; Blood Platelets; Buffers; Cells; Characteristics; Charge; Clinic; Clinical; DNA; Detection; Devices; Direct Costs; Dyes; Escherichia coli; Escherichia coli K12; Flow Cytometry; Fluorescence; Fluorescent Probes; Funding; Goals; Human; Image; Incubated; Individual; Label; Leukocytes; Life; Lighting; Marketing; Measurement; Measures; Medicine; Methodology; Methods; Monitor; Performance; Phase; Protocols documentation; Reagent; Real-Time Systems; Research; Risk; Sampling; Schedule; Scheme; Scientist; Screening procedure; Simulate; Small Business Technology Transfer Research; Solutions; Staining method; Stains; Stream; System; Techniques; Technology; Testing; Time; Transfusion; United States; Universities; Vascular blood supply; Whole Blood; Wyoming; base; blood product; cost; design; digital; innovation; microorganism; pathogenic bacteria; prototype; public health relevance; rapid detection; research and development

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.

描述(由申请人提供)：这个STTR第一阶段项目将证明一种低成本、快速检测浓缩血小板中细菌污染的系统的概念，只需几分钟而不是几天，性价比与公认的测试方法相当。尽管美国拥有世界上最安全的血液供应之一，但输注受致病菌污染的血液产品存在重大危险。在美国，细菌感染是除人为错误之外最危险的输血相关风险。血库将血小板培养培养2天以进行细菌检测。由于血小板的保质期为5天，保质期缩短了2天，对供应造成了严重影响。建议的解决方案将使血小板寿命延长30%以上，并能够释放比旧血小板更安全、更有效的新鲜血小板。这项拟议的研发工作的目标是建立和测试一个概念验证系统，并开发一种检测浓缩血小板单位中细菌的染色方法。拟议的专利方法被称为喷泉流“细胞计数法”。这种方法是基于这样一种系统，在该系统中，用LED照明含有用荧光探针标记的微生物的溶液流，并用数码相机检测荧光。与其他设备相比，低成本商用数码相机和LED的使用使这种设备在经济上具有吸引力。这项拟议的研究将用一个系统测试替代染料组合，该系统旨在检测红细胞和白细胞背景下加入到分离血小板中的细菌。预期的结果是替代更昂贵、更不敏感和更慢的检测系统。喷泉流动“将允许在自动培养设备的灵敏度下检测到血小板污染，但只需约10分钟。该系统可以自动化，每天可能允许检查数百个血小板单位。将对接种到人类采集器中的血小板单位的大肠杆菌进行测试。关键的性能标准将是对大肠杆菌的方法检测限(MDL)、总检测时间、检测效率和假阳性检测率。我们的总体性能目标是证明该技术的商业可行性，MDL为~1细菌/毫升(在10毫升样本中)，检测效率为&gt；75%(检测到细菌/细菌存在)，流速为&gt；3.0毫升/分钟。这个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 Inc.的一组科学家与其怀俄明大学STTR合作伙伴、一名输血医学顾问(丹佛邦菲尔斯血液中心)和一名临床微生物学家(克利夫兰诊所)共同进行。与公共卫生相关：在美国，每年大约有400万个单位的浓缩血小板被输注，估计每1000到3000个单位中就有一个被潜在的致病菌污染。被细菌污染的血液现在是除了人为错误之外，美国最危险的输血相关风险。该项目提出了一种系统，该系统将提供更低的成本，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。