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）：使用核酸扩增测试技术（NAT）最近开发了商业上可用的测定法，使血液中心有可能考虑将这些测试应用于献血者筛查。但是，到目前为止，NAT测试系统是耗时的，需要受限制的实验室空间和受过良好训练的技术人员，并且通常不适合大规模筛查供体样品。美国血液中心采用了两种基本的“集合和池解决方案”策略。一个是24的直池，其中“反应池”的分辨率需要测试单个样品。在另一种策略中，准备较小的（中级）池并组合以创建最终（主）池。反应性主池的分辨率是首先测试中间池，然后在必要时进行单个样品。池大小和集合策略平衡测定敏感性与确保血液供应的实际需求。 Maxwell Sensors，Inc。（MSI）建议将依赖性酶依赖酶的扩增（HDA）和数字磁珠（DM-Bead）技术组合，以在高吞吐量的96粒微型板格式中生成一个样品快速，同时进行多种病原体（SAMP-B）的同时测定。在单个微孔中，几滴供体的血液与DM珠混合在一起，可以同时识别HIV 1和2型HIV，乙型肝炎（HBV），丙型肝炎（HCV），人类T-淋巴细胞性病毒（HTLV I型和II）（HTLV I型和II）（HTLV I型），West Nile Virus（West Nile Virus），West Virus（WNV）和许多其他胰岛症。 DM珠，由光刻造影制成的数字条形码，然后用核酸探针标记，能够准确地识别单个均匀培养基中的许多靶标。 合并的技术非常强大，并提供以下优势： 不需要样本池：可以在个人的样本上对多种病原体进行筛查。 少量样品：鉴定多种靶病原体所需的全部需要几滴血液。它不仅决定了一个反应性样本，而且还标识了“反应性”。 减少窗口周期：HDA提供了高灵敏度和特异性，而没有与基于抗体的筛选技术相关的“窗口周期”。 快速和高通量：可以在单个96孔微板板上执行简单的等温手术和96个患者样品。 灵活性：轻松添加新的探针，以在珠子上进行其他筛选目标。 低成本，易于使用和高可靠性：DM珠子是低成本，简单的操作步骤，批次变化是最小的。 在此项目期间，我们将重点关注四个关键任务：1）数字磁条形码珠的设计和制造，2）设计和构建检测系统，3）测试基于DM-BEAD的多重HDA测定和4）评估系统和生物测定性能。公共卫生相关性：Maxwell Sensors Inc.（MSI）建议使用数字磁性条形码珠开发基于解旋酶的基于放大的系统，这些珠为供体血液的小型标本中的多种血源性病原体提供快速，准确且易于使用的筛查。这种对血液中多种病原体的同时测定将有助于血库和临床实验室快速测试大量捐赠的血液，并有助于防止血传播感染传播。提出的技术将晚期半导体制造过程与分子特征放大和探测结合，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。