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）：此STTR I期项目将证明低成本快速系统的概念，用于在几分钟而不是几天内，成本/性能等于公认的测试方法，用于检测血小板浓缩物中的细菌污染。尽管美国拥有世界上最安全的血液供应之一，但由于被致病细菌污染的血液产物的输血而存在着重大危险。细菌感染代表了除人类错误以外，美国最危险的输血相关风险。血库将血小板培养物孵育2天进行细菌测试。由于血小板单元的保质期为5天，因此保质期2天的缩短对供应有严重影响。所提出的溶液将使血小板寿命延长30％以上，并可以释放比较旧血小板更安全，更有效的新鲜血小板。这项提出的研发工作的目的是建立和测试概念验证系统，并开发一种用于检测血小板浓缩单位细菌的染色方法。所提出的专家方法称为喷泉流量“细胞术”。该方法基于一个系统，在该系统中，包含用荧光探针标记的微生物的溶液流被LED照明，并通过数码相机检测到荧光，并通过数字摄像头检测到荧光。在使用该设备上，该设备可以使该设备的使用在ADEN上进行替代的替代方法。检测细菌在红细胞和白色血细胞的背景下尖刺到平台上。该系统可以自动化，并有可能每天检查数百个血小板单元。测试将对接种到人体形态学血小板单元中的大肠杆菌进行测试。关键性能标准将是大肠杆菌，总测定时间，检测效率和假阳性检测率的方法检测极限（MDL）。我们证明该技术商业生存能力的总体性能目标是〜1个细菌/mL的MDL（在10mL样品中），检测效率> 75％（检测到的细菌/细菌），流速为3.0 mL/分钟。该项目的关键任务是开发将实现这些目标的染色试剂组合。这项研究将由Softray Inc.的科学家团队与其怀俄明大学的STTR合作伙伴，输血医学顾问（Bonfils Blood Center，Denver）和临床微生物学家（Cleveland Clinic）进行合作。公共卫生相关性：每年在美国输血大约400万单位的血小板浓缩物，估计有1000万至1千分之一的三千单位被潜在的病原细菌污染。被细菌污染的血液现在代表了美国除人类错误以外的其他与美国相关的风险。该项目提出了一个系统，该系统将提供对细菌污染的较低成本，更及时，更敏感的检测，从而为输血提供更安全的血小板供应。