Sensitive Detection of Bacterial Contamination in Platelets

描述（由申请人提供）：血小板细菌污染是输血传播感染中发病率和死亡率的主要原因。据估计，血小板的细菌污染率为1 / 2000至1 / 3000血小板单位。根据监测和其他研究的推断，这些输血造成的预计死亡人数从每年15人到200多人不等。目前有几种检测血小板细菌的技术方法，但由于缺乏所需的速度或灵敏度，没有一种被广泛接受。Lynntech提出了一种创新的、易于操作的、定量的、实时对流PCR （QC-PCR）热循环细菌检测系统（BDS），以实现血小板在释放时间附近检测所需的速度和灵敏度。基于QC-PCR的BDS从核酸提取到扩增、实时检测，在1小时内实现随机核酸检测。该技术预计能够检测血小板中广泛的细菌，检测限为102至103 CFU /mL（每次PCR反应1至10个基因组拷贝）。由于血小板，特别是汇集的随机供者全血来源的血小板，在释放前立即进行处理，因此需要快速的细菌检测来检测离散样本。因此，该系统的随机获取核酸分析能力满足了输血服务的关键需求，因为输血服务不能同时处理大量的血小板。用户不需要等待其他PCR反应完成后再运行更多的样品。这种创新的系统将有助于减少处理和释放血小板单位的延迟，目前与基于培养的方法相关。此外，每次检测只需要200升的血小板。这将显著减少以培养为基础的方法的血小板损失，每种方法需要3至5ml的血小板。简而言之，该系统的实施将显著减少感染病例和死亡人数，提高患者的生活质量，并为医疗保健系统每年节省数百万美元。此外，重要的是系统的简单性，这使得医院和血库可以购买低成本的系统。Lynntech基于QC-PCR的BDS是下一代分子诊断工具，它将使非专业操作人员能够在任何环境下以更快的速度和更低的成本进行复杂的生物测试。该QC-PCR技术可以适应运行几乎任何现有的和未来的PCR为基础的临床诊断分析。通过大大简化和加速检测，北斗系统可以在医生办公室和医院以即时医疗形式进行分子诊断检测。该体系对资源贫乏的国家也大有裨益。这项技术为更好地管理血小板供应质量提供了机会，每年影响数百万美国人，特别是预防因输血污染血小板而引起的细菌相关感染和败血症。考虑到市场规模和通过延长血小板保质期和改善感染预防而节省的相关费用，经济影响也是巨大的。一旦开发出来，该产品还可以扩展用于检测其他血液制品成分中的病原体。此外，该系统将具有巨大的商业潜力和医疗价值，可在医生办公室、诊所、医院甚至资源贫乏的国家提供快速、低成本、易于使用的体外诊断。

DESCRIPTION (provided by applicant): Bacterial contamination of platelets is the leading cause of morbidity and mortality from a transfusion- transmitted infection. Bacterial contamination rates for platelets have been estimated to occur at a frequency of 1 in 2,000 to 1 in 3,000 platelet units. Projected fatalities from these transfusions, based on extrapolation from surveillance and other studies, vary from 15 to over 200 per year. There are several technical approaches for bacterial detection in platelets but none have been widely accepted because they either lack the speed or sensitivity required. Lynntech proposes an innovative, easy-to-operate, quantitative, real-time convective PCR (QC-PCR) thermal cycling bacteria detection system (BDS) to achieve the speed and sensitivity required for platelets testing near the time of release. The QC-PCR based BDS provides random-access nucleic acid testing in one hour, from nucleic acid extraction to amplification and real-time detection. The technology is anticipated to be able to detect a broad range of bacteria in platelets at a detection limit between 102 to 103 CFU per/mL (1 to 10 genomic copies per PCR reaction). Because platelets, especially the pooled random donor whole-blood derived platelets, are processed immediately before release, rapid bacterial detection is needed to test the discrete samples. The system's random-access nucleic acid analysis capability, therefore, meets the critical needs of transfusion services which do not process high numbers of platelets for transfusion simultaneously. Users will not need to wait for completion of other PCR reactions before running more samples. This innovative system will help reduce the delay in processing and release of platelet units that are currently associated with culture based methods. In addition, each test requires only 200 ¿L of platelets. This will significantly reduce the loss of platelets to culture-based methods which each require 3 to 5 mL of platelets. In short, the implementation of the system will significantly reduce the number of cases of infections and fatalities, improving the patients' quality of life and saving the health care system millions of dollars annually. Also, important is the simplicity of the system which leads to low-cost systems for hospitals and blood banks to purchase. Lynntech's QC-PCR based BDS is the next generation of tool for molecular diagnostics that will enable a non-specialized operator to perform sophisticated biological tests in any setting with greater speed at a lower cost. The QC-PCR technology can be adapted to run virtually any existing and future PCR based clinical diagnostic assays. By significantly simplifying and accelerating testing, the BDS system could allow for molecular diagnostic testing at doctors' offices as well as hospitals in point-of-care format. The system will also be great for resource poor countries. This technology provides an opportunity for better management of the quality of platelet supply affecting millions of Americans annually, specifically for preventing bacterial related infections and sepsis through transfusion of contaminated platelets. The economic impact is also enormous considering the market size and associated savings from prolonging platelet shelf life and improving infection prevention. Once developed, the product can also be expanded for use to detect pathogens in other blood product components. Also, the system will have great commercial potential and medical values in providing rapid, low-cost, easy-to-use in- vitro diagnostics in doctors' offices, clinics, hospitals, and even in countries with poor resources.