SBIR PA22-176 - Culture-free microbial enrichment for diagnosis and characterization of anti-microbial resistance

SBIR PA22-176 - 用于诊断和表征抗微生物耐药性的免培养微生物富集

• 批准号: 10699317
• 负责人: David F Allison
• 金额: $ 30万
• 依托单位: TRIANGLE BIOTECHNOLOGY, INC.
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-30 至 2024-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10699317
• 关键词: SBIR; PA22; 176; Culture; free

Abstract Timely identification of infectious agents enables targeted antibiotic administration and better health outcomes for patients. However, current methodologies take days to months, leading to the administration of broad- spectrum antibiotics when they may not be appropriate or effective. Drastic cost reductions of DNA sequencing technologies have led to increased demand for such workflows towards the identification of infectious agents. However, clinical samples often have extremely low levels of pathogens present. Additionally, depending upon the particular microorganism, the genetic payload is not always easy to access due to lysis-resilient cell wall structures. Finally, there is often overwhelming contamination from non-target DNA derived from the host or benign microorganisms that are part of the healthy microbiome. This project will establish the feasibility of a novel targeted-microbubble approach, Levios™, to enrich infectious pathogens directly from clinical samples and enable comprehensive diagnostics and strain identification for fast and personalized antimicrobial treatments. Enrichment of target microbes will enable the success of culture-free, molecular-based diagnostics via targeted enrichment of pathogenic microbial organisms. Triangle’s unique approach to microbial enrichment will address the clinical need for higher diagnostic sensitivity using direct clinical samples. In this Phase I proposal, we will demonstrate proof-of-concept of our Levios enrichment technology using Candida auris antimicrobial resistance (AMR) characterization as a real-world application for culture-free molecular-based assays. This is an ideal candidate to display the advantages of Levios as C. auris is an emerging multi-drug resistant pathogenic threat in the United States, is resilient to lysis, can live on hard surfaces for extended periods of time, and can colonize the skin. We will use our Levios microbial enrichment reagent to concentrate and enrich C. auris from swabs and blood samples to increase the sensitivity of detection for culture-free, molecular-based diagnostics. In alignment with our objectives, we will also develop digital PCR (dPCR)-based assays to identify the presence of pathogenic Candida strains (including C. auris) in patient samples and will include antimicrobial resistance information. We will also develop genomic sequencing workflows to characterize the strain of infectious Candida, which will provide more detailed strain information for epidemiology and AMR gene tracking. The proposed work will demonstrate that the Levios technology is capable of capturing target microbes from human samples, resulting in concentrated, high-quality samples for culture-free molecular-based diagnostics. The results will support commercialization of the platform in Phase II.

抽象的 及时识别传染源可以实现有针对性的抗生素管理和更好的健康结果 对于患者。然而，当前的方法需要数天到数月的时间，导致广泛的管理 当谱抗生素可能不合适或无效时。 DNA 测序成本大幅降低 技术导致对识别传染原的工作流程的需求增加。 然而，临床样本中的病原体含量通常极低。此外，还取决于 对于特定的微生物，由于细胞壁具有裂解弹性，遗传有效负载并不总是容易获得 结构。最后，常常存在来自宿主或宿主的非目标 DNA 的大量污染。 良性微生物是健康微生物组的一部分。该项目将建立一个可行性 新型靶向微泡方法 Levios™，可直接从临床样本中富集传染性病原体 并实现全面的诊断和菌株鉴定，以实现快速和个性化的抗菌 治疗。目标微生物的富集将使无培养、基于分子的诊断取得成功 通过有针对性地富集病原微生物。 Triangle 独特的微生物富集方法 将满足使用直接临床样本进行更高诊断灵敏度的临床需求。在这个第一阶段 提案中，我们将使用耳念珠菌展示 Levios 富集技术的概念验证 抗菌素耐药性 (AMR) 表征作为免培养分子基于的实际应用 化验。这是展示 Levios 优势的理想候选者，因为耳念珠菌是一种新兴的多药药物 在美国具有抗病原威胁，具有抗裂解能力，可以在硬表面上长时间生存 随着时间的推移，可以在皮肤上定植。我们将使用我们的 Levios 微生物富集试剂来浓缩和富集 来自拭子和血液样本的耳念珠菌，可提高无培养、基于分子的检测灵敏度 诊断。为了实现我们的目标，我们还将开发基于数字 PCR (dPCR) 的检测方法来识别 患者样本中存在致病性念珠菌菌株（包括耳念珠菌），并将包括抗菌药物 电阻信息。我们还将开发基因组测序工作流程来表征菌株 传染性念珠菌，将为流行病学和 AMR 基因追踪提供更详细的菌株信息。 拟议的工作将证明 Levios 技术能够捕获目标微生物 人类样本，产生浓缩的高质量样本，用于无培养的分子诊断。 结果将支持该平台第二阶段的商业化。