Culture and amplification-free bacterial sepsis diagnosis

无培养和扩增细菌败血症诊断

• 批准号: 10484211
• 负责人: Alfredo Andres Celedon
• 金额: $ 29.86万
• 依托单位: SCANOGEN, INC.
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-01 至 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10484211
• 关键词: Achievement; Architecture; Bacteremia; Bacteria; Biological Assay; Blood; Clinical; Clinical Research; Complication; Detection; Development; Device or Instrument Development; Diagnosis; Diagnostic; Engineering; Goals; Head; Hospitals; Hour; Infection; Length of Stay; Life; Manuals; Methods; Microbiology; Molecular; Organism; Patient-Focused Outcomes; Patients; Performance; Phase; Physicians; Pilot Projects; Preparation; Procedures; Process; Protocols documentation; Publishing; Reagent; Recommendation; Resources; Ribosomal RNA; Sampling; Sensitivity and Specificity; Sepsis; Species Specificity; Specificity; System; Techniques; Technology; Testing; Time; United States Food and Drug Administration; Universities; Whole Blood; Work; antimicrobial; assay development; base; cost effective; detection limit; detection method; fungus; improved; instrument; manufacturability; microorganism; mortality; multidisciplinary; multiplex assay; novel; operation; pathogen; protocol development; rapid detection; rapid test; septic patients; single molecule; targeted treatment; usability; validation studies

Culture and amplification-free bacterial sepsis diagnosis Summary Sepsis is a life-threatening complication of an infection that requires immediate treatment. The identification of the causative microorganism utilizing processes based on blood culture is a keystone of sepsis diagnosis. However, blood culture takes one to five days to provide results. Here, we propose the development of a culture and amplification-free assay to detect and identify bacteria in one hour directly on whole blood. The new assay will enable physicians to provide therapy targeting the causative organism days earlier than the current standards. The new assay will be based on Single MOLecule Tethering (SMOLT), a novel sample preparation and amplification-free molecular detection method developed at Scanogen. The work proposed in this application will build on the achievements of an ongoing project aimed at developing an assay to detect fungi in whole blood. The SMOLT-Bacteria assay proposed in this application will identify the species, genus or order of the causative organism in approximately 90% of bloodstream infections. The new system will be easy-to-use, fully automated, and capable of rapid detection in whole blood without blood culture. Despite the urgent clinical need, there is only one FDA approved culture-free molecular assay for sepsis diagnosis. In comparison to this test, the SMOLT- Bacteria assay will be 4 times faster, 6 times less expensive, and thanks to its higher multiplexing capability, it will detect and identify the causative pathogen in twice the number of patients. In our preliminary results, we developed a multiplex assay capable of detecting bacteria directly in whole blood. We demonstrated that the assay is highly specific and sensitive with a limit of detection (LOD) between 1-10 CFU/mL, depending on the species, which is as good as the only FDA-approved test. In this project, we will complete the development of the assay protocol by including new probes and optimizing the overall assay (Aim 1). We will test the manual protocol and develop a fully integrated disposable cartridge and instrument (Aim 2). Finally, we will evaluate the assay in pilot analytical and clinical validation studies (Aim 3). 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 Food and Drug Administration (FDA). If successful, the new automated assay can become part of the standard procedure for the diagnosis of patients with sepsis and improve patients’ outcomes by dramatically reducing the time to targeted antimicrobial therapy.

无培养和扩增细菌败血症诊断 概括 脓毒症是一种危及生命的感染并发症，需要立即治疗。 利用基于血培养的过程来确定致病微生物是脓毒症诊断的关键。 然而，血培养需要一到五天才能提供结果，在这里，我们建议开发一种培养物。 以及无需扩增的检测方法，可在一小时内直接对全血进行细菌检测和鉴定。 将使医生能够比目前的治疗提前几天针对致病微生物提供治疗 标准。 新的测定将基于单分子束缚（SMOLT），这是一种新颖的样品制备和 Scanogen 开发的无扩增分子检测方法。本申请中提出的工作。 将建立在一个正在进行的项目所取得的成就的基础上，该项目旨在开发一种检测全血中真菌的方法。 本申请中提出的 SMOLT-细菌测定将鉴定致病菌的种、属或目 大约 90% 的血流感染中都有微生物。新系统将易于使用、完全自动化、 尽管临床需求迫切，但能够在全血中快速检测，无需进行血培养。 与该测试相比，只有一种 FDA 批准的用于脓毒症诊断的无培养分子测定法。 细菌检测速度将提高 4 倍，成本降低 6 倍，并且由于其更高的多重检测能力， 将使两倍数量的患者检测并识别致病病原体。 在我们的初步结果中，我们开发了一种能够直接检测全血中细菌的多重检测方法。 我们证明该测定具有高度特异性和敏感性，检测限 (LOD) 在 1-10 之间 CFU/mL，取决于物种，这与 FDA 批准的唯一测试一样好。 通过包含新探针和优化整体测定来完成测定方案的开发（目标 1) 我们将测试手动方案并开发完全集成的一次性试剂盒和仪器（目标 2）。 最后，我们将在试点分析和临床验证研究中评估该测定（目标 3）。 多学科团队，包括分析开发和仪器开发方面的专家，包括 Becton Dickinson 前工程副总裁，微生物学、败血症诊断和 来自约翰·霍普金斯大学和匹兹堡大学的治疗，以及来自 Key 的工程师 技术。完成这个项目后，我们将进一步优化系统的可用性和可制造性。 以及提交给食品和药物管理局 (FDA) 的分析行为和临床研究。 成功后，新的自动化检测可以成为患者诊断标准程序的一部分 败血症并通过显着减少靶向抗菌治疗的时间来改善患者的治疗结果。