Point-of-care detection of TB and NTM pathogens with Fluorescent Deoxyribozyme Sensors and 3D-printed, battery powered device.

使用荧光脱氧核酶传感器和 3D 打印电池供电设备对 TB 和 NTM 病原体进行即时检测。

• 批准号: 9889371
• 负责人: Yulia Gerasimova
• 金额: $ 14.53万
• 依托单位: UNIVERSITY OF CENTRAL FLORIDA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-01-22 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9889371
• 关键词: 3D Print; Address; Antibiotics; Bacillus; Bacteria; Bedside Testings; Binding; Biological Assay; Brazil; Case Study; Catalytic DNA; Cessation of life; Child; Chronic Disease; Cleaved cell; Clinical; Collaborations; Color; Communicable Diseases; Complementarity Determining Regions; Complex; Country; Cystic Fibrosis; DNA; Data; Detection; Devices; Diagnosis; Diagnostic; Disease; Disease Outcome; Disease Progression; Evaluation; Failure; Fluorescence; Freeze Drying; Genus Mycobacterium; Goals; Gold; HIV; HIV Seropositivity; In Vitro; Incidence; Infectious Diseases Research; Infrastructure; Injectable; International; Kinetics; Label; Lung; Lung infections; Methods; Microscopy; Molecular; Morbidity - disease rate; Mycobacterium Infections; Mycobacterium tuberculosis; Nanotechnology; Nucleic Acid Amplification Tests; Nucleic Acids; Oral; Outcome; Pathogenicity; Patients; Performance; Pharmaceutical Preparations; Physicians; Polymerase; Population; Public Health; RNA; Reagent; Refractory; Resources; Sampling; Sensitivity and Specificity; Signal Transduction; Site; Specificity; Speed; Sputum; Technology; Testing; Time; Treatment Protocols; Tube; Tuberculosis; Validation; bacterial lysate; base; clinically relevant; co-infection; cohort; comorbidity; cost; diagnostic assay; effective therapy; flexibility; fluorophore; global health; improved; innovation; instrument; mortality; mycobacterial; non-tuberculosis mycobacteria; novel; pathogen; point of care; point-of-care diagnostics; portability; prototype; rRNA Genes; recombinase; sensor; tool; transmission process; tuberculosis diagnostics

Project Summary Tuberculosis (TB), an infectious disease caused by species of Mycobacterium tuberculosis complex (MTC), remains one of the major public health problems worldwide, with ~10 million new cases and about ~1.5 million deaths each year. With ~70,000 new cases reported in 2017, Brazil remains among the 22 high-burden countries that account for >80% of all TB cases worldwide. Non-tuberculous mycobacteria (NTM) are an emerging group of related opportunistic pathogens that cause TB-like pulmonary infections, particularly in patients with underlying comorbidities such as HIV or cystic fibrosis. NTM infections are notoriously refractory to treatment, with <50% cure rates for some species despite year-long treatment regimens with antibiotic cocktails of oral and injectable drugs. This situation is further exacerbated by HIV co-infection which promotes disease progression and complicates diagnosis of TB and NTM pulmonary infections. An estimated 3.5 million cases of TB go undetected each year, largely due to the lack of affordable, effective point-of-care diagnostics for TB/NTM infection. As a result, infected patients develop more severe disease and continue to transmit these pathogens. The frequent misdiagnosis of NTM infections as TB by the commonly used smear microscopy method delays appropriate treatment which can result in worse clinical outcomes. Thus, point-of-care (POC) tests are urgently needed to provide physicians with actionable data required to effectively treat patients with these debilitating chronic diseases. The goal of this project is to take advantage of recent advancements in DNA nanotechnology, molecular sensors, and 3D-printing to significantly improve POC diagnostics capability for TB and NTM. Our platform combines the speed and sensitivity of RPA isothermal amplification, the specificity of binary deoxyribozyme (BiDz) sensors, and economy and portability of a 3D-printed assay device. We will optimize multi-color multiplex sensor assays for specific detection of TB and NTM pathogens from sputum samples. A cheap, portable battery-powered device for assay incubation and fluorescent detection built using 3D-printing technology will be tested. Finally, “real-world” validation of this diagnostic assay will be conducted in Brazil on samples from HIV+ and HIV- cohorts. If successful, this flexible technology could be exploited to devise POC diagnostic assay for other infectious diseases. An added outcome of this international collaboration will be enhancement of the infectious disease research capacity and infrastructure in Brazil.

项目概要 结核病 (TB) 是一种由结核分枝杆菌复合体 (MTC) 物种引起的传染病， 仍然是全球主要公共卫生问题之一，新增病例约 1000 万，新增病例约 150 万 每年都有死亡人数。 2017 年报告新增病例约 7 万例，巴西仍是 22 个高负担国家之一 占全球所有结核病病例的 80% 以上。非结核分枝杆菌（NTM）是一个新兴群体 引起结核样肺部感染的相关机会病原体，尤其是患有潜在结核病的患者 HIV或囊性纤维化等合并症。众所周知，NTM 感染很难治疗，<50% 尽管使用口服和注射抗生素混合物进行长达一年的治疗方案，但某些物种的治愈率 药物。 HIV 合并感染进一步加剧了这种情况，从而促进疾病进展和 使 TB 和 NTM 肺部感染的诊断变得复杂。估计有 350 万结核病病例未被发现 每年，这主要是由于缺乏负担得起的、有效的结核病/非结核分枝杆菌感染即时诊断方法。作为一个 结果，受感染的患者会发展出更严重的疾病并继续传播这些病原体。频繁的 常用涂片镜检方法将 NTM 感染误诊为结核病，延误了适当的治疗时间 治疗可能会导致更差的临床结果。因此，迫切需要进行即时护理（POC）测试 为医生提供有效治疗这些使人衰弱的慢性病患者所需的可操作数据 疾病。该项目的目标是利用 DNA 纳米技术的最新进展， 分子传感器和 3D 打印可显着提高 TB 和 NTM 的 POC 诊断能力。我们的 平台结合了 RPA 等温扩增的速度和灵敏度、二元分析的特异性 脱氧核酶 (BiDz) 传感器，以及 3D 打印检测装置的经济性和便携性。我们会优化 多色多重传感器检测，用于特异性检测痰样本中的 TB 和 NTM 病原体。一个 使用 3D 打印构建的廉价便携式电池供电设备，用于测定孵化和荧光检测 技术将受到考验。最后，该诊断测定的“现实世界”验证将于 2019 年在巴西进行 来自 HIV+ 和 HIV- 队列的样本。如果成功，这种灵活的技术可用于设计 POC 其他传染病的诊断测定。此次国际合作的另一个成果将是 加强巴西的传染病研究能力和基础设施。