Highly Multiplexed Single Molecule Tethering

高度多重化的单分子束缚

• 批准号: 10760792
• 负责人: Alfredo Andres Celedon
• 金额: $ 100万
• 依托单位: SCANOGEN, INC.
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-17 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10760792
• 关键词: Antibiotics; Architecture; Area; Biological Assay; Blood; Blood Circulation; Blood capillaries; Blood specimen; Clinical; Clinical Research; Computer software; Cytolysis; Detection; Development; Device or Instrument Development; Diagnosis; Diagnostic tests; Engineering; Etiology; Future; Goals; Head; Hospitals; Image; Manuals; Mass Spectrum Analysis; Methods; Microbe; Microbiology; Molecular; Optics; Organism; Pathogen detection; Patients; Performance; Pilot Projects; Preparation; Price; Procedures; Process; Protocols documentation; Reagent; Resources; Risk; Sampling; Sensitivity and Specificity; Sepsis; Specificity; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Symptoms; System; Technology; Testing; Time; Universities; Whole Blood; Work; antimicrobial; assay development; commercialization; cost effective; cross reactivity; design; detection assay; detection limit; detection method; diagnostic assay; instrument; manufacturability; manufacture; microbial; microorganism; mortality; multidisciplinary; multiplex assay; novel; novel strategies; operation; pathogen; prototype; single molecule; standard of care; usability; validation studies

Highly Multiplexed Single Molecule Tethering Summary Bloodstream infections in patients with sepsis symptoms should be promptly treated with antibiotics; however, the time required to identify pathogens using methods based on blood culture is a major obstacle to timely effective antimicrobial therapy. The development of a comprehensive molecular assay capable of identifying microbes directly from whole blood has been elusive despite major efforts due to the low concentration of microbes in the bloodstream and the large number of microbial species that cause bloodstream infection (200+). We have developed an automated assay that detects 13 bloodstream microbes directly in whole blood at the low concentrations found in clinical samples, with LODs between 0.5 and 3 CFU/mL and a turnaround time of 70 minutes. The assay performance is superior to other FDA cleared assays for bloodstream pathogen detection. The new assay is based on Single MOLecule Tethering (SMOLT), a novel sample preparation and amplification-free molecular detection technology developed at Scanogen. Here, we propose to dramatically increase the multiplexing capacity of SMOLT using a novel approach with the goal of producing a panel that covers the organisms responsible for more than 99% of bloodstream infection cases (Aim 1). We will also integrate the automated assay into a single disposable cartridge that will be processed by an easy-to-use and fully automated desktop instrument (Aim 2). We will evaluate the new assay in pilot clinical studies where we will directly compare the performance of the new assay to the standard of care diagnostic test based on blood culture followed by mass spectroscopy (Aim 3). Each of these specific aims has quantitative milestones. Our long-term goal is the development of an FDA-cleared and CLIA-waived system; this project is a critical step towards that goal. We will address the challenges of achieving the required level multiplexing and assay integration, which are the most serious risks on the path to developing the clinical system. We will work with a multidisciplinary team that includes experts in assay development and instrument development, including the former Vice President of Engineering at Becton Dickinson, experts in microbiology, sepsis diagnosis and treatment from Johns Hopkins University and the University of Pittsburgh, as well as engineers from Key Technologies. After completing this project, we will further optimize the system for usability and manufacturability and conduct analytical and clinical studies for submission to the FDA. If successful, the new automated assay can become part of the standard procedure for the diagnosis of patients suspected of having sepsis. The new highly multiplexed assay will be useful in other applications where rapid, cost-effective, and highly multiplexed molecular detection is needed.

高度多重的单分子束缚 概括 脓毒症症状患者的血液感染应及时治疗 抗生素；但是，使用基于血液的方法识别病原体所需的时间 培养是及时有效抗菌治疗的主要障碍。一个发展 能够直接从全血中鉴定的微生物的综合分子测定 由于血液中的微生物浓度较低而导致了巨大的努力，但仍难以捉摸 以及引起血液感染的大量微生物物种（200+）。我们有 开发了一种自动化测定 在临床样品中发现的低浓度，LOD在0.5至3 CFU/mL之间 周转时间为70分钟。测定性能优于其他FDA清除测定法 用于血液病原体检测。新测定基于单分子链接 （Smolt），一种新型样品制备和无扩增的分子检测技术 在扫描原开发。在这里，我们建议大幅度提高 Smolt使用一种新颖的方法，目的是生产覆盖生物体的面板 负责超过99％的血液感染病例（AIM 1）。我们还将整合 自动化测定到单个一次性墨盒中，该弹药筒将通过易于使用和 全自动桌面仪器（AIM 2）。我们将评估试点临床研究中的新测定 我们将直接将新测定的性能与护理标准进行比较 基于血液培养的诊断测试，然后是质谱法（AIM 3）。每个 具体目标具有定量的里程碑。 我们的长期目标是开发FDA清理和Clia捕获的系统。这个项目 是朝着这个目标迈出的关键一步。我们将解决达到所需水平的挑战 多路复用和测定集成，这是发展道路上最严重的风险 临床系统。我们将与包括分析专家在内的多学科团队合作 开发和仪器开发，包括前工程副总裁 Becton Dickinson，Johns Hopkins的微生物学专家，败血症诊断和治疗 大学和匹兹堡大学以及关键技术的工程师。后 完成该项目，我们将进一步优化系统，以实现可用性和可制造性和 进行分析和临床研究以提交FDA。如果成功，新的 自动化测定可以成为患者诊断标准程序的一部分 怀疑患有败血症。新的高度多路复用测定将在其他应用中有用 需要快速，成本效益和高度多重分子检测。