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: 8315698
• 负责人: Gao Chen
• 金额: $ 49.79万
• 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 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8315698
• 关键词: Antibodies; Automation; Bar Codes; Biological Assay; Blood; Blood Banks; Blood Screening; Blood Tests; Blood donor; Blood donor screening; Blood-Borne Pathogens; Chemistry; Clinical; Computer software; Coupled; Detection; Development; Diagnostic; Disease; Donor Selection; Drops; Evaluation; Fluorescence; Genes; Genetic Materials; Goals; HIV; HIV-1; Hepatitis B; Hepatitis B Virus; Hepatitis C; Hepatitis C virus; Hour; Human; Human Resources; Human T-lymphotropic virus 1; Individual; Infection; Infectious Agent; Label; Laboratories; Lead; Magnetic Bead Technology; Magnetism; Methodology; Methods; Molecular; Molecular Profiling; Nucleic Acid Amplification Tests; Nucleic Acids; One-Step dentin bonding system; Performance; Phase; Phycoerythrin; Procedures; Process; Reaction; Reproducibility; Research; Research Project Grants; Resources; Running; Sampling; Screening procedure; Semiconductors; Sensitivity and Specificity; Specificity; Specimen; Streptavidin; Syphilis; System; Technology; Testing; Time; Training; United States; Variant; Virus; West Nile virus; analog; base; cost; design; digital; flexibility; helicase; instrument; magnetic beads; operation; optical imaging; pathogen; prevent; product development; prototype; sensor; verification and validation

ABSTRACT Rapid, high-throughput, and highly accurate identification of potential blood borne diseases will help blood banks to quickly test a very large volume of donated blood, and will help to prevent further infections. Current blood testing has the following issues: 1. Time consuming testing and sample operation that must be performed for all diseases "one-by-one". 2. A large sample volume is required that depends on the number of tests. 3. Testing procedure is complicated as the number of tests increase. 4. Complicated operational procedures that must be run by highly trained personnel in molecular laboratories. 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, not in "multiplexed (> 2) fashion", thus are unsuited to large-scale screening of donor samples for various pathogens. Maxwell Sensors, Inc. (MSI) proposes to combine Helicase-Dependent Amplification and Barcode Magnetic Bead (BMB) technologies to produce a rapid, Simultaneous Assay for Multiple Pathogens in Blood (SAMP-B). With a few drops of a donor's blood combined with digital magnetic beads in a single microwell, it is possible to simultaneously identify HIV type 1 and 2, Hepatitis B (HBV), Syphilis, Hepatitis C (HCV), Human T-Lymphotropic Virus (HTLV type I and II), West Nile Virus (WNV), and many other blood-borne diseases. During this Phase I project, we have completed all tasks and successfully demonstrated the technical feasibility of simultaneous assay for multiple pathogens by fabricating 128-plex BMBs, integrating the analyzer, and performing HIV, HBV, and Syphilis multiplexed assays. The resulting combined technology is very powerful and will offer the following advantages: (1) 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" (2) Reduced window period: offers high sensitivity and specificity without the "window period" associated with antibody based screening technologies (3) Rapid and high throughput: the system can automatically process up to 100 samples per hour - for a maximum of 12,800 reportable results in a 96-microwell format (4) Flexibility: easy addition of new probes for additional screening targets on beads (5) Low cost, easy to use, and high reliability: BMB are low cost, simple operation steps, and batch to batch variation is minimal. In this Phase II project, we will focus on: (1) expand blood-borne pathogens panels from (3-plex to 8-plex) for Blood screening, (2) integrate the sample process with SAMP-B analyzer, and (3) develop and evaluate multiplex pathogens for blood screening.

抽象的 快速、高通量、高度准确地识别潜在的血源性疾病将有助于血库快速检测大量的捐献血液，并有助于防止进一步的感染。目前的血液检测存在以下问题： 1. 必须对所有疾病“一一”进行耗时的检测和取样操作。 2. 需要大量样本，具体取决于测试数量。 3.随着测试次数的增加，测试程序变得复杂。 4. 操作程序复杂，必须由经过严格培训的人员在分子实验室进行操作。 最近使用核酸扩增测试技术（NAT）的商业化检测的发展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 测试系统是时间密集型的，不是“多重（> 2）方式”，因此不适合大规模筛查供体样本的各种病原体。 Maxwell Sensors, Inc. (MSI) 提议将解旋酶依赖性扩增和条形码磁珠 (BMB) 技术结合起来，对血液中的多种病原体进行快速同步检测 (SAMP-B)。通过将几滴献血者的血液与单个微孔中的数字磁珠结合，可以同时识别 HIV 1 型和 2 型、乙型肝炎 (HBV)、梅毒、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。在这一阶段的项目中，我们完成了所有任务，并通过制造 128 重 BMB、集成分析仪以及进行 HIV、HBV 和梅毒多重检测，成功证明了同时检测多种病原体的技术可行性。由此产生的组合技术非常强大，并将具有以下优势： （1）样本量小：微孔中仅需几滴血液即可鉴定多种目标病原体。它 不仅确定反应样本，还识别“对什么反应” (2) 缩短窗口期：提供高灵敏度和特异性，没有与基于抗体的筛选技术相关的“窗口期” (3) 快速、高通量：系统每小时可自动处理多达 100 个样品 - 96 微孔格式最多可报告 12,800 个结果 (4) 灵活性：轻松添加新探针，以在珠子上添加额外的筛选目标 (5)成本低、使用方便、可靠性高：BMB成本低、操作步骤简单、批次间偏差极小。 在此第二阶段项目中，我们将重点关注：(1) 将用于血液筛查的血源性病原体组合从（3 重至 8 重）扩展，(2) 将样本处理与 SAMP-B 分析仪集成，以及 (3) 开发和评估用于血液筛查的多重病原体。