Rapid and automated detection of bloodborne pathogens for improved treatment and antimicrobial stewardship

快速自动检测血源性病原体，以改善治疗和抗菌管理

• 批准号: 10546973
• 负责人: Lorenzo Damico
• 金额: $ 30.27万
• 依托单位: AINCOBIO LLC
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10546973
• 关键词: Address; Animal Model; Antibodies; Antimicrobial susceptibility; Bacteria; Bacterial Infections; Biological Assay; Blood; Blood Circulation; Blood specimen; Blood-Borne Pathogens; Centrifugation; Clinical; Clinical Microbiology; Clinical Trials; Colony-forming units; Comparative Study; DNA; DNA sequencing; Data; Density Gradient Centrifugation; Detection; Devices; Diagnostic; Diagnostic tests; Distant; Economics; Escherichia coli; Foundations; Genome; Goals; Growth; Health; Healthcare Systems; Hospitals; Hour; Immune response; In Vitro; Infection; Lab-On-A-Chips; Label; Laboratories; Libraries; Life; Logistics; Measures; Metabolic; Methods; Microbe; Microbiology; Molecular; Molecular Analysis; Mycoses; Organism; Pathogen detection; Patients; Pharmaceutical Preparations; Phase; Phenotype; Predisposition; Preparation; Primary Infection; Process; Recovery; Risk; Sampling; Science; Sensitivity and Specificity; Sepsis; Services; Site; Small Business Innovation Research Grant; Spectrum Analysis; System; Testing; Time; Tube; Validation; Whole Blood; antimicrobial; antimicrobial drug; clinical diagnostics; clinically relevant; cost; density; design; detection limit; drug resistant microorganism; effective therapy; electric impedance; fluid flow; fungus; implementation facilitation; improved; in-vitro diagnostics; individualized medicine; instrument; low and middle-income countries; microbial; miniaturize; molecular diagnostics; mortality risk; nanopore; pathogen; pathogenic bacteria; pathogenic fungus; phase 2 study; prototype; targeted treatment; treatment planning

PROJECT SUMMARY Sepsis is a life-threatening condition caused by bacterial infection of the bloodstream that threatens nearly two million lives in the U.S. annually. Risk of death escalates quickly once bacteria or fungi enter the bloodstream; thus, quick and accurate detection of bloodborne pathogens is critical to inform immediate tailored treatment. However, the current standard practice to detect bloodborne bacteria and fungi requires in vitro culture to measure microbial growth, a process which takes 12-120 hours, with times to a confirmed negative sometimes taking as long as 5 days. Positive cultures then undergo additional tests to determine bacterial identity and antimicrobial susceptibility. This protracted process, which includes significant pre-analytical delays caused by sample transport to off-site microbiology labs and long culture times, represents a critical bottleneck in the diagnostic laboratory workflow, creating significant economic and labor burden to the healthcare system and prolonging the time that patients may be receiving inappropriate treatment (broad spectrum antimicrobial drugs instead of targeted therapy). Aincobio is developing a rapid, near patient in vitro diagnostic test, the BactoPhore, that can be deployed outside of the centralized lab to isolate and detect intact and viable low-concentration bacteria and fungi in blood specimens in 2 hours. BactoPhore achieves enhanced analytical sensitivity by rapidly concentrating pathogens for detection to deliver a positive result quickly. To do this, BactoPhore incorporates a “lab-on-a-chip” system that can isolate microbes from 0.5 mL whole blood using dielectrophoresis (DEP) and detect metabolic signatures using electrochemical impedance spectroscopy (EIS) within a low-cost consumable. A diagnostic prototype of the BactoPhore isolates >90% of target bacteria in 30 minutes and can detect 100 CFUs of E. coli within 2 hours. Further, target microbes pre-concentrated from 10 mL of whole blood can be trapped by DEP in <30 minutes, demonstrating the potential for processing clinically relevant volumes. In this Phase I SBIR, Aincobio will demonstrate feasibility of commercializing the BactoPhore test for label-free, antibody-independent, and culture-independent detection, concentration, and isolation of low abundance and viable bacterial and fungal pathogens by 1) evaluating the integrated BactoPhore prototype with common pathogens spiked into whole blood, and 2) demonstrating that BactoPhore enables rapid and sensitive downstream sequencing of the target microbe by depleting host DNA prior to extraction and library preparation. Completion of these aims will establish proof-of-concept that BactoPhore has analytical sensitivity to confirm infection from whole blood within 2 hours and significantly improve time-to-results for the entire diagnostic workflow from positive result to pathogen identification.

项目摘要 脓毒症是一种危及生命的条件所造成的细菌感染的血液，威胁近两个 每年有100万人在美国生活。一旦细菌或真菌进入血液，死亡风险迅速上升; 因此，快速和准确地检测血液传播的病原体对于提供及时的有针对性的治疗至关重要。 然而，目前检测血液传播细菌和真菌的标准实践需要体外培养， 测量微生物生长，这一过程需要12-120小时，有时需要时间才能确认阴性 耗时长达5天。阳性培养物然后进行额外的测试以确定细菌身份， 抗菌敏感性这一旷日持久的过程包括分析前的重大拖延， 样品运输到非现场微生物实验室和长培养时间，代表了 诊断实验室工作流程，对医疗保健系统造成重大的经济和劳动负担， 延长患者可能接受不适当治疗（广谱抗菌药物）的时间 而不是靶向治疗）。Aincobio正在开发一种快速、接近患者的体外诊断测试，BactoPhore， 可以部署在中心实验室之外，以隔离和检测完整的和可行的低浓度 血液样本中的细菌和真菌。BactoPhore通过快速的 浓缩病原体进行检测，以快速提供阳性结果。为此，BactoPhore结合了 “芯片实验室”系统，可以使用介电泳（DEP）从0.5 mL全血中分离微生物， 使用电化学阻抗谱（EIS）检测低成本消耗品内的代谢特征。 BactoPhore的诊断原型在30分钟内分离>90%的目标细菌，并且可以检测100 E.大肠杆菌在2小时内。此外，可以将从10 mL全血中预浓缩的靶微生物 在<30分钟内被DEP捕获，证明了处理临床相关体积的潜力。在这 第一阶段SBIR，Aincobio将证明商业化BactoPhore测试的可行性， 抗体非依赖性和培养非依赖性的检测、浓缩和分离低丰度和 活的细菌和真菌病原体，通过1）评估具有常见细菌和真菌病原体的整合的BactoPhore原型， 病原体掺入全血，和2）证明BactoPhore能够快速和灵敏地 通过在提取和文库制备之前耗尽宿主DNA对靶微生物进行下游测序。 这些目标的完成将确立BactoPhore具有分析灵敏度的概念验证，以确认 在2小时内从全血中感染，并显著改善整个诊断的结果时间 从阳性结果到病原体鉴定的工作流程。