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

项目总结 脓毒症是一种危及生命的疾病，由血液中的细菌感染引起，威胁到近两个 每年有数百万人生活在美国。一旦细菌或真菌进入血液，死亡风险就会迅速上升； 因此，快速准确地检测血液传播的病原体对于立即进行量身定制的治疗至关重要。 然而，目前检测血液传播细菌和真菌的标准做法需要体外培养 测量微生物生长，这是一个需要12-120小时的过程，有时会出现确认的阴性 耗时长达5天。然后，阳性培养物接受额外的测试，以确定细菌鉴定和 抗菌药物敏感性。这一旷日持久的过程，包括由以下原因造成的重大分析前延迟 将样本运送到异地微生物实验室和较长的培养时间，是 诊断实验室工作流程，给医疗保健系统造成了巨大的经济和劳动力负担 延长患者可能接受不适当治疗的时间(广谱抗菌药物 而不是靶向治疗)。Aincobio正在开发一种快速的、接近患者的体外诊断测试，即BactoPhore， 可以部署在集中式实验室之外，以分离和检测完整且可行的低浓度 血标本中细菌和真菌在2小时内。BactoPhore通过快速提高分析灵敏度 集中病原体进行检测，以迅速产生阳性结果。要做到这一点，BactoPhore结合了一个 “芯片实验室”系统，可使用介电泳法(DEP)从0.5毫升全血中分离微生物 在低成本消耗品中使用电化学阻抗谱(EIS)检测代谢特征。 一种诊断原型的BactoPhore在30分钟内分离出90%的目标细菌，并可以检测到100 在2小时内检测到大肠杆菌的CFU。此外，从10毫升全血中预浓缩的目标微生物可以 在&lt；30分钟内被DEP捕获，展示了处理临床相关卷的潜力。在这 第一阶段SBIR，Aincobio将展示将无标签的BactoPhore测试商业化的可行性， 抗体非依赖和培养独立的检测、浓缩和分离低丰度和 活的细菌和真菌病原体通过1)评估整合的BactoPhore原型与普通 病原体大量进入全血，以及2)证明了BactoPhore能够快速和敏感地 在提取和文库准备之前，通过消耗宿主DNA对目标微生物进行下游测序。 这些目标的完成将建立概念验证，即BactoPhore具有分析敏感度以确认 在2小时内从全血感染，并显著提高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。

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.