Simultaneous Assay for Multiple Pathogens in Blood (SAMP-B)

血液中多种病原体的同时检测 (SAMP-B)

• 批准号: 7744078
• 负责人: Gao Chen
• 金额: $ 20万
• 依托单位: MAXWELL SENSORS, INC.
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-03 至 2010-08-02
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7744078
• 关键词: Adopted; Antibodies; Automation; Bar Codes; Beds; Biological Assay; Birds; Blood; Blood Banks; Blood donor; Blood donor screening; Blood-Borne Pathogens; Clinical; Detection; Development; Diagnostic; Disease; Donor Selection; Drops; Equilibrium; Evaluation; Figs - dietary; Fluorescence; Goals; HIV; Hepatitis B; Hepatitis B Virus; Hepatitis C; Hepatitis C virus; Human; Individual; Infection; Laboratories; Magnetic Bead Technology; Magnetism; Molecular; Molecular Profiling; Nucleic Acid Amplification Tests; Nucleic Acid Probes; Operative Surgical Procedures; Patients; Performance; Persons; Polymers; Process; Property; Protocols documentation; Reaction; Resolution; Sampling; Screening procedure; Semiconductors; Sensitivity and Specificity; Specimen; Syphilis; System; Technology; Testing; Tetanus Helper Peptide; Time; Training; Variant; Viral; Virus; West Nile virus; base; cost; design; design and construction; digital; flexibility; fluorophore; helicase; indium arsenide; magnetic beads; pathogen; prevent; public health relevance; sensor

DESCRIPTION (provided by applicant): The recent development of commercially-available assays using nucleic acid amplification test technologies (NAT) have made it possible for blood centers to consider applying these tests to blood donor screening. But NAT test systems, so far, are time intensive, require restricted laboratory space and highly-trained technical staff, and generally are unsuited to large-scale screening of donor samples. U.S. blood centers have adopted two basic "pooling and pool resolution" strategies. One is a straight pool of 24 in which resolution of a "reactive pool" requires testing individual samples. In the other strategy, smaller (intermediate) pools are prepared and combined to create a final (master) pool. Resolution of reactive master pools occurs by first testing the intermediate pools and then, if necessary, the individual samples. Pool size and pooling strategy balance assay sensitivity with the practical demands of assuring blood availability. Maxwell Sensors, Inc. (MSI) proposes to combine Helicase-Dependent Amplification (HDA) and Digital Magnetic Bead (DM-Bead) technologies to produce a one sample rapid, Simultaneous Assay for Multiple Pathogens in Blood (SAMP-B), in a high throughput 96-well microplate format. With a few drops of a donor's blood combined with DM-beads in a single microwell, it is possible to simultaneously identify HIV type 1 and 2, Hepatitis B (HBV), Hepatitis C (HCV), Human T- Lymphotropic Virus (HTLV type I and II), West Nile Virus (WNV), and many other blood-borne diseases. The DM-Bead, mass fabricated with digital bar codes by photolithography, then tagged with nucleic acid probes, are able to accurately identify many targets in a single homogeneous medium. The combined technology is very powerful and offers the following advantages: Sample pooling is not necessary: screening for multiple pathogens can be performed on an individual person's sample. Small quantity of sample: a few drops of blood in a microwell are all that's needed to identify multiple target pathogens. It not only determines a reactive sample, but also identifies "reactive to what". Reduced window period: HDA offers high sensitivity and specificity without the "window period" associated with antibody based screening technologies. Rapid and high throughput: simple isothermal operation and 96 patient samples can be performed on a single 96-well microplate. Flexibility: easy addition of new probes for additional screening targets on beads. Low cost, easy to use, and high reliability: DM-beads are low cost, simple operation steps, and batch to batch variation is minimal. During this project, we will focus on four key tasks: 1) design and fabrication of digital magnetic barcode beads, 2) design and construction of a detection system, 3) testing the DM-Bead-based multiplex HDA assay and 4) evaluation of system and bioassay performance. PUBLIC HEALTH RELEVANCE: Maxwell Sensors Inc. (MSI) proposes to develop a helicase-dependent amplification-based system, using digital magnetic barcoded beads that provide rapid, accurate, and easy-to-use screening for multiple blood-borne pathogens in small specimens of donor blood. This Simultaneous Assay for Multiple Pathogens in Blood will help blood banks and clinical laboratories to quickly test very large volumes of donated blood, and will help prevent the spread of blood-borne infections. The proposed technology that marries an advanced semiconductor fabrication process with molecular signature amplification and probing will allow high-throughput molecular diagnostic profiling of individuals at relatively low cost.

描述（由申请人提供）：最近开发的使用核酸扩增测试技术（NAT）的商用化验使得血液中心可以考虑将这些测试应用于献血者筛查。但迄今为止，NAT 测试系统时间密集，需要有限的实验室空间和训练有素的技术人员，并且通常不适合大规模筛选供体样本。美国血液中心采用了两种基本的“汇集和汇集解决”策略。一种是 24 个样本的直接池，其中“反应池”的分辨率需要测试单个样本。在另一种策略中，准备并组合较小的（中间）池以创建最终（主）池。解决反应性主池的方法是首先测试中间池，然后在必要时测试各个样本。池大小和池策略平衡测定灵敏度与确保血液可用性的实际需求。 Maxwell Sensors, Inc. (MSI) 提议将解旋酶依赖性扩增 (HDA) 和数字磁珠 (DM-Bead) 技术相结合，以高通量 96 孔微孔板形式生产单样本快速同时检测血液中多种病原体 (SAMP-B)。通过在单个微孔中加入几滴捐赠者的血液与 DM-bead 结合，可以同时识别 HIV 1 型和 2 型、乙型肝炎 (HBV)、丙型肝炎 (HCV)、人类 T 淋巴细胞病毒（HTLV I 型和 II 型）、西尼罗河病毒 (WNV) 和许多其他血液传播疾病。 DM-Bead 通过光刻技术用数字条形码大规模制造，然后用核酸探针标记，能够准确识别单一均质介质中的许多目标。 组合技术非常强大，并具有以下优点： 不需要合并样本：可以对个人样本进行多种病原体筛查。 样品量小：只需微孔中的几滴血液即可识别多种目标病原体。它不仅确定反应样本，还识别“对什么反应”。 缩短窗口期：HDA 具有高灵敏度和特异性，没有与基于抗体的筛选技术相关的“窗口期”。 快速、高通量：简单的等温操作和 96 个患者样本可以在单个 96 孔微孔板上进行。 灵活性：轻松添加新探针，以在珠子上进行额外的筛选目标。 成本低、使用方便、可靠性高：DM-bead成本低、操作步骤简单、批次间差异极小。 在这个项目中，我们将重点关注四项关键任务：1）数字磁性条形码珠的设计和制造，2）检测系统的设计和构建，3）测试基于DM-Bead的多重HDA测定以及4）系统和生物测定性能的评估。公共健康相关性：Maxwell Sensors Inc. (MSI) 提议开发一种基于解旋酶依赖性扩增的系统，该系统使用数字磁条形码珠，可以快速、准确且易于使用地筛查供体血液小样本中的多种血源性病原体。这种对血液中多种病原体的同时检测将帮助血库和临床实验室快速检测大量捐献的血液，并有助于防止血源性感染的传播。所提出的技术将先进的半导体制造工艺与分子特征放大和探测相结合，将允许以相对较低的成本对个体进行高通量分子诊断分析。