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.

培养和无扩增细菌败血症诊断 概括 败血症是需要立即治疗的感染的生命并发症。识别 利用基于血液培养过程的过程的病因微生物是败血症诊断的键石。 但是，血液培养需要一到五天才能提供结果。在这里，我们建议一种文化的发展 和无扩增的测定，以直接在全血上直接在一小时内检测和鉴定细菌。新测定 将使医生能够比目前的日期更早地提供针对因果生物的治疗 标准。 新测定将基于单分子绑扎（Smolt），一种新型样品制备和 在扫描原开发的无扩增分子检测方法。本申请中提出的工作 将基于一个正在进行的项目的成就，旨在开发一种检测全血真菌的测定法。 本应用中提出的Smolt-细菌测定法将确定因果关系的物种，属或顺序 大约90％的血液感染中有机体。新系统将易于使用，完全自动化， 并且能够在没有血液培养的情况下快速检测。尽管紧急临床需要，但仍有 仅FDA批准了无文化的分子测定法，用于败血症诊断。与此测试相比，Smolt- 细菌测定法的速度将快4倍，价格便宜6倍，并且要感谢其更高的多重能力 将检测和识别两倍患者数量的因果病原体。 在我们的初步结果中，我们开发了一种能够直接在全血中检测细菌的多重测定法。 我们证明该测定具有高度特异性和敏感性，并且在1-10之间的检测极限（LOD） CFU/ML，取决于物种，这与唯一的FDA批准测试一样好。在这个项目中，我们将 通过包括新问题并优化整体测定法（AIM 1）。我们将测试手动协议，并开发完全集成的一次性弹药筒和仪器（AIM 2）。 最后，我们将评估试验分析和临床验证研究中的测定法（AIM 3）。我们将与 多学科团队，包括分析开发和仪器开发专家，包括 Becton Dickinson的前工程副总裁，微生物学专家，败血症诊断和 约翰·霍普金斯大学和匹兹堡大学的治疗，以及Key的工程师 技术。完成该项目后，我们将进一步优化系统的可用性和制造性 并进行分析和临床研究，以提交食品药物管理局（FDA）。如果 成功，新的自动化测定可以成为患者诊断的标准程序的一部分 通过败血症并改善患者的结局，从而大大减少了靶向抗菌治疗的时间。