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 及时识别传染剂可以实现目标抗生素给药和更好的健康结果 适用于患者。但是，当前的方法需要几天到几个月，从而导致了广泛的管理 频谱抗生素可能不合适或有效。 DNA测序的急剧成本降低 技术导致对此类工作流程的需求增加，以识别传染剂。 但是，临床样品通常存在极低的病原体。另外，取决于 特定的微生物，由于裂解弹性细胞壁，基因有效载荷并不总是容易获得的 结构。最后，从宿主派生的非目标DNA或 良性微生物是健康微生物组的一部分。该项目将确定 新型的靶向微泡方法Levios™直接从临床样品中富集感染性病原体 并为快速和个性化的抗菌剂启用全面的诊断和应变识别 治疗。目标微生物的富集将使基于培养的，基于分子的诊断能够成功 通过靶向富集致病微生物的富集。三角形的独特微生物富集方法 将使用直接临床样本来满足更高诊断敏感性的临床需求。在这个阶段第一 提案，我们将使用Candida Auris证明我们的LEVIOS富集技术的概念证明 抗菌耐药性（AMR）表征作为无培养分子的现实应用 测定。这是显示Levios的优势的理想候选人，因为C. Auris是新兴的多药 美国的抗致病威胁对裂解具有抗性，可以在硬表面上生存长时间 时间，可以定居皮肤。我们将使用LEVIOS微生物富集试剂进行浓缩和浓缩 拭子和血液样品中的弧菌，以提高对无培养，基于分子的检测的敏感性 诊断。为了与我们的目标保持一致，我们还将开发基于数字PCR（DPCR）的测定法以识别 在患者样品中存在致病性念珠菌菌株（包括Auris），并将包括抗菌素 阻力信息。我们还将开发基因组测序工作流程，以表征 感染性念珠菌将为流行病学和AMR基因跟踪提供更详细的菌株信息。 拟议的工作将证明LEVIOS技术能够从中捕获目标微生物 人类样品，导致基于无培养的分子诊断的浓缩，高质量的样品。 结果将支持该平台在第二阶段的商业化。