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(申请人提供)：使用核酸扩增检测技术(NAT)的商业可用检测技术的最新发展使血液中心考虑将这些检测应用于献血者筛查。但到目前为止，NAT测试系统是时间密集型的，需要有限的实验室空间和训练有素的技术人员，而且通常不适合大规模筛选捐赠者样本。美国血液中心采用了两种基本的“池和池解决”策略。一种是由24个人组成的直接池，在这个池中，“反应池”的解析需要测试单独的样品。在另一种策略中，准备较小的(中间)池并将其合并以创建最终的(主)池。反应主池的解决方法是首先测试中间池，然后如有必要，再测试单个样本。池的大小和池策略平衡了检测灵敏度和确保血液可用性的实际需求。 Maxwell Sensors，Inc.(MSI)建议将解旋酶依赖放大(HDA)和数字磁珠(DM-Bead)技术相结合，以高通量96孔微孔板格式生产一种血液中多种病原体的快速、同时分析(SAMP-B)样品。只需在单个微孔中滴几滴献血者的血液和DM珠子，就有可能同时识别HIV 1型和2型、乙肝(乙肝)、丙型肝炎(丙型)、人类T淋巴细胞病毒(HTLV I和II)、西尼罗河病毒(WNV)和许多其他血液传播疾病。DM-Bead通过光刻技术大量制造数字条形码，然后标记核酸探针，能够在单一均匀介质中准确识别多个目标。 组合后的技术非常强大，并提供以下优势： 样本池不是必须的：可以对单个人的样本进行多种病原体的筛查。 少量样本：只需在微孔中滴几滴血就可以识别多个目标病原体。它不仅确定一个反应性样本，而且还确定“对什么反应”。 缩短窗口期：HDA提供高灵敏度和特异度，无需与基于抗体的筛查技术相关的“窗口期”。 快速和高通量：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：易于添加新的探头，以增加珠子上的筛查目标。 成本低，使用方便，可靠性高：DM珠子成本低，操作步骤简单，批次之间的差异最小。 在该项目中，我们将重点完成四项关键工作：1)数字条形码磁珠的设计和制造，2)检测系统的设计和构建，3)基于DM-Bead的多路HDA检测，4)系统和生物检测性能的评估。与公共健康相关：Maxwell Sensors Inc.(MSI)建议开发一种基于解旋酶的扩增系统，使用数字磁性条形码珠子提供快速、准确和易于使用的筛查小样本献血者血液中的多种血液传播病原体。这种同时检测血液中多种病原体的方法将有助于血库和临床实验室快速检测大量捐献的血液，并有助于防止血液传播感染的传播。这项拟议的技术将先进的半导体制造工艺与分子签名放大和探测结合在一起，将允许以相对较低的成本高通量地对个体进行分子诊断。