AstraDx: Sub-Hour AST Via Computational Image Processing

AstraDx：通过计算图像处理实现不到一小时的 AST

• 批准号: 10547198
• 负责人: Jonathan Florez
• 金额: $ 29.96万
• 依托单位: ASTRADX, INC.
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10547198
• 关键词: Acinetobacter baumannii; Address; Agreement; American; Antibiotics; Antimicrobial Resistance; Antimicrobial susceptibility; Area; Bacteria; Behavior; Biotechnology; Blood; Boston; Caring; Categories; Centers for Disease Control and Prevention (U.S.); Cessation of life; Code; Communicable Diseases; Complex; Dangerousness; Data; Data Set; Detection; Devices; Disease; Drug Combinations; ESKAPE pathogens; Engineering; Ensure; Enterococcus; Eye; Fostering; Genus staphylococcus; Goals; Gold; Growth; Guidelines; Health; Home; Hospitals; Hour; Human; Image; Incubators; Infection; Intelligence; Lead; Legal patent; Life; Measures; Mechanics; Methods; Microbiology; Minimum Inhibitory Concentration measurement; Mission; Modernization; Morbidity - disease rate; Optics; Organism; Outcome; Patients; Performance; Pharmaceutical Preparations; Phase; Phenotype; Process; Pseudomonas aeruginosa; Public Health; Resistance; Resolution; Resources; Running; Scientist; Sepsis; Side; Small Business Innovation Research Grant; Sum; Technology; Testing; Time; Training; United States National Institutes of Health; Validation; Vancomycin resistant enterococcus; Work; biosafety level 2 facility; carbapenem resistance; computational pipelines; cost; disease diagnostic; drug standard; experimental study; image processing; improved; innovation; member; methicillin resistant Staphylococcus aureus; mortality; mortality risk; multidrug-resistant Pseudomonas aeruginosa; normal microbiota; optical imaging; pathogen; personalized diagnostics; prevent; priority pathogen; routine Bacterial stain; serial imaging

PROJECT SUMMARY/ABSTRACT Antimicrobial susceptibility testing (AST) for bacterial bloodstream infections (BSIs) is dangerously slow. Starting from a positive blood culture, AST currently takes a median of 15 hours, and often takes days. Until results are available, there is no choice but to treat patients with empiric antibiotics and hope the pathogen is susceptible. This is unacceptable. BSIs are serious, rapidly fatal infections that kill a quarter-million Americans each year, including a third of all hospital deaths. Empiric antibiotics destroy patients’ normal flora, predisposing them to additional, untreatable infections. Every hour a patient is on the wrong antibiotic increases risk of death by 7.6%. This key problem has made rapid, affordable AST a global priority. To address this problem, AstraDx has developed a low-cost new AST device that we hypothesize can perform AST in under an hour. The purpose of this proposal is to test this hypothesis. Preliminary studies involving multiple experiments on 19 strains vs. 8 standard antibiotics have demonstrated growth detection in 31±18 minutes (95% range, 20-63 minutes). Strains tested included methicillin-resistant S. aureus (MRSA), vancomycin-resistant Enterococci (VRE), carbapenem-resistant A. baumannii (CRAB), carbapenem-resistant Enterobacterales (CRE), multidrug-resistant P. aeruginosa—all high priority per the CDC—as well as corresponding susceptible strains. The innovation that makes these results possible is a new computational image-processing pipeline, which enables extremely sensitive and robust detection of an antibiotic’s effect on growth, often in less than the strain’s doubling time. A second innovation is the use of inexpensive standard components, leading to an affordable device and consumables. These innovations support our long-term goal of making rapid AST universally available. We will test our hypothesis through three tightly focused specific aims. Aim 1 will generate training data on diverse bug-drug combinations, with bacteria from simulated- positive blood cultures grown in the presence of doubling dilutions of antibiotic on 384-well plates, similar to the current phenotypic gold standard, with multiple antibiotics on each plate. Aim 2 will use the images from Aim 1 to create a unified picture of each strain’s behavior in the presence of each antibiotic, which we will use to refine how we determine minimum inhibitory concentrations (MICs) in order to maximize categorical agreement with the CLSI gold standard. Finally, Aim 3 will validate the method from Aim 2 by testing additional bacterial strains and measuring categorical agreement with the gold standard, per FDA standard guidelines. The expected outcome is a device proven to achieve sub-hour AST with FDA-level performance on a wide variety of pathogens, establishing technical merit, feasibility, and commercial potential for an SBIR Phase II. Our advanced technology, a clearly identified product, will help meet the critical need for rapid, highly sensitive infectious-disease diagnostics to address antimicrobial resistance for life-threatening BSIs and sepsis.

项目概要/摘要 针对细菌性血流感染 (BSI) 的抗菌药物敏感性测试 (AST) 速度极其缓慢。 目前，从血培养呈阳性开始，AST 检测平均需要 15 小时，通常需要数天。直到 结果出来了，别无选择，只能用经验性抗生素治疗患者，并希望病原体被消灭 易受影响的。这是不可接受的。 BSI 是严重且迅速致命的感染，导致 25 万美国人死亡 每年，其中包括三分之一的医院死亡人数。经验性抗生素会破坏患者的正常菌群， 使他们容易遭受额外的、无法治疗的感染。每小时都有一名患者服用错误的抗生素 死亡风险增加 7.6%。这一关键问题使得快速、经济的 AST 成为全球优先事项。 为了解决这个问题，AstraDx 开发了一种低成本的新型 AST 设备，我们假设它可以执行 不到一小时即可获得 AST。该提案的目的是检验这一假设。初步研究涉及 对 19 种菌株与 8 种标准抗生素进行的多次实验已证明在 31±18 中检测到生长 分钟（95% 范围，20-63 分钟）。测试的菌株包括耐甲氧西林金黄色葡萄球菌 (MRSA)、 耐万古霉素肠球菌 (VRE)、耐碳青霉烯鲍曼不动杆菌 (CRAB)、耐碳青霉烯类抗生素 肠杆菌 (CRE)、多重耐药铜绿假单胞菌 — CDC 均高度重视 — 以及 相应的敏感菌株。使这些结果成为可能的创新是一种新的计算 图像处理管道，能够极其灵敏和稳健地检测抗生素对人体的影响 生长，通常在不到菌株倍增的时间内。第二个创新是使用廉价的标准 组件，从而提供价格实惠的设备和消耗品。这些创新支持我们的长期目标 使快速 AST 普遍可用。我们将通过三个紧密关注的具体内容来检验我们的假设 目标。目标 1 将生成有关各种错误药物组合的训练数据，其中细菌来自模拟- 阳性血培养物在 384 孔板上加倍稀释的抗生素下生长，类似于 目前的表型金标准，每个板上有多种抗生素。目标 2 将使用目标 1 中的图像 创建每个菌株在每种抗生素存在下的行为的统一图片，我们将用它来 完善我们确定最低抑制浓度 (MIC) 的方法，以最大限度地提高分类一致性 符合 CLSI 黄金标准。最后，目标 3 将通过测试额外的细菌来验证目标 2 的方法 根据 FDA 标准指南，菌株和测量与金标准绝对一致。 预期结果是一种设备被证明能够在广泛的应用中实现亚小时 AST 并具有 FDA 级别的性能 各种病原体，确定 SBIR 第二阶段的技术优点、可行性和商业潜力。 我们的先进技术和明确标识的产品将有助于满足快速、高灵敏度的关键需求 传染病诊断，以解决危及生命的 BSI 和败血症的抗菌药物耐药性问题。