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.

摘要 及时识别传染源可实现有针对性的抗生素管理和更好的健康结果 对患者然而，目前的方法需要几天到几个月的时间，导致广泛的管理， 当抗生素可能不合适或无效时。DNA测序成本大幅降低 技术的发展导致对鉴定传染原的这种工作流程的需求增加。 然而，临床样品通常具有极低水平的病原体存在。此外，根据 对于特定的微生物，由于细胞壁具有裂解弹性， 结构.最后，通常存在来自宿主或宿主细胞的非靶DNA的压倒性污染。 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 一种新的靶向微泡方法，Levios™，用于直接从临床样本中富集感染性病原体 实现全面的诊断和菌株鉴定， 治疗。目标微生物的富集将使无培养、基于分子的诊断获得成功 通过有针对性地富集病原微生物。三角洲独特的微生物富集方法 将解决使用直接临床样品的更高诊断灵敏度的临床需求。这项I期 在一项提案中，我们将使用耳念珠菌证明我们的Levios富集技术的概念验证 抗菌素耐药性（AMR）表征作为一个现实世界的应用，无培养的分子为基础的 测定。这是一个理想的候选人，以显示Levios作为C的优势。奥瑞司是一种新兴的多药 在美国，抗病原体威胁，对溶解有弹性，可以在硬表面上长时间存活 时间，可以殖民皮肤。我们将使用我们的Levios微生物富集试剂进行浓缩和富集 C.耳拭子和血液样本，以增加检测的灵敏度，无培养，分子为基础的 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， 致病性念珠菌菌株（包括C.耳），并将包括抗微生物剂 阻力信息我们还将开发基因组测序工作流程，以表征 传染性念珠菌，这将提供更详细的流行病学和AMR基因跟踪菌株信息。 这项拟议的工作将证明Levios技术能够捕获来自 人类样本，从而产生浓缩的高质量样本，用于无培养的分子诊断。 研究结果将支持该平台在第二阶段的商业化。