A highly multiplexed convection flow qPCR assay and platform delivering rapid, comprehensive identification and predictive AST of pathogens causing UTIs.

高度多重对流 qPCR 检测和平台，可对引起 UTI 的病原体进行快速、全面的识别和预测 AST。

• 批准号: 10322806
• 负责人: Dmitriy Khodakov
• 金额: $ 30万
• 依托单位: TORUS BIOSYSTEMS, INC.
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10322806
• 关键词: Accounting; Affect; Antibiotic Resistance; Antibiotic Therapy; Antibiotics; Antimicrobial Resistance; Bacteria; Biological Assay; Biomedical Engineering; Cells; Chemistry; Clinical; Clinical Sensitivity; Collaborations; Community Hospitals; Complex; Convection; Cytolysis; DNA; Detection; Development; Diagnosis; Diagnostic; Diagnostic tests; Drug resistance; Fractionation; Future; Genes; Genomics; Gold; Health Care Costs; Healthcare; Infection; Infection Control; Laboratories; Medical; Molecular Diagnostic Techniques; Morbidity - disease rate; Mutation; Nature; Nosocomial Infections; Nucleic Acid Amplification Tests; Nucleic Acids; Nucleotides; Paper; Pathogen detection; Patient-Focused Outcomes; Patients; Performance; Phase; Population; Preparation; Prevalence; Rapid diagnostics; Reaction; Recovery; Recurrence; Reproducibility; Sampling; Sensitivity and Specificity; Small Business Innovation Research Grant; Solid; Specimen; System; Technology; Testing; Therapeutic; Time; TimeLine; Urinary tract infection; Urine; Uropathogen; Validation; Visit; Work; amplification detection; antimicrobial; base; clinical application; clinical research site; communicable disease diagnosis; design; detection sensitivity; diagnostic assay; diagnostic panel; diagnostic platform; improved; insight; molecular diagnostics; multi-drug resistant pathogen; pathogen; pathogenic bacteria; pathogenic fungus; personalized medicine; point of care; point-of-care diagnostics; prototype; rapid test; resistance gene; synthetic construct

Project Summary Urinary tract infections (UTIs) are among the most prevalent community- and hospital-acquired infections (~10M annual medical visits), accounting for considerable morbidity and annual US healthcare spend of greater than $6B. While UTI pathogens are well understood, patients are frequently treated empirically. The lack of rapid, point of care molecular diagnostics that can identify UTI pathogens results in poor antimicrobial stewardship and high rates of UTI drug resistance. We propose to begin development of a rapid diagnostic assay and workflow that will concurrently detect all UTI pathogens and their associated resistance genes, suitable for point of care clinical applications. This assay is anticipated to allow for more effective personalized treatment of UTIs, reducing rates of antimicrobial resistance, and improving patient outcomes. As described in an upcoming Nature Biomedical Engineering paper, the Torus Biosystems’s qPCR system, Synestia™, has demonstrated reproducible 30-plex capability, accurate quantitation and SNP detection, sensitivity down to 10 genomic copies, and turnaround times of less than 30 minutes. The consumable is equipped with an embedded pre-quenched microarray that allows for real-time detection of amplification products without opening the reaction chamber. The sensitivity and dynamic range of the Synestia system has been evaluated across a range of input concentrations of DNA (101 to 105 copies) and the threshold value was found to be dependent on the log of the input DNA concentration. The platform has also successfully demonstrated rapid bacteria identification using a multiplex panel specific for 15 bacteria species. This Phase 1 application proposes to develop a 60-plex UTI panel for comprehensive assessment of uropathogen DNA within 30 minutes on the Synestia platform. In the subsequent Phase 2, we will develop, optimize, and validate a fully integrated consumable including urine sample prep (lysis and extraction) on the Synestia platform and validate clinical performance in collaboration with select clinical sites. The team anticipates that the demonstrated assay will constitute a solid basis for further development of a commercial point of care diagnostic test for urinary tract infections.

项目概要 尿路感染 (UTI) 是最常见的社区和医院获得性感染之一 （每年约 1000 万次就诊），占美国每年医疗保健支出的相当大比例 大于 $6B 虽然 UTI 病原体已被充分了解，但患者经常接受经验治疗。 缺乏能够识别尿路感染病原体的快速护理点分子诊断导致治疗效果不佳 抗菌药物管理和尿路感染耐药率高。 我们建议开始开发快速诊断测定和工作流程，以同时检测 所有 UTI 病原体及其相关耐药基因，适合护理点临床应用。 该检测预计将能够对尿路感染进行更有效的个性化治疗，降低尿路感染发生率 抗菌素耐药性，并改善患者的治疗效果。 正如即将发表的《自然生物医学工程》论文中所述，Torus Biosystems 的 qPCR 系统， Synestia™ 已展示可重复的 30 重能力、准确的定量和 SNP 检测， 灵敏度低至 10 个基因组拷贝，周转时间少于 30 分钟。 配备嵌入式预猝灭微阵列，可实时检测扩增 无需打开反应室即可获得 Synestia 系统的灵敏度和动态范围。 已对一系列 DNA 输入浓度（101 至 105 个拷贝）和阈值进行了评估 该平台还发现该值取决于输入 DNA 浓度的对数。 成功展示了使用针对 15 种细菌的多重检测板的快速细菌鉴定 物种。 该第一阶段申请建议开发一个 60 重 UTI 面板，用于全面评估 在 Synestia 平台上 30 分钟内检测尿路病原体 DNA 在接下来的第 2 阶段，我们将开发， 优化并验证完全集成的耗材，包括尿液样本制备（裂解和提取） Synestia 平台并与选定的临床中心团队合作验证临床表现。 预计所证实的测定将为进一步开发 尿路感染的商业护理点诊断测试。