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: 10321573
• 负责人: Yulia Gerasimova
• 金额: $ 14.74万
• 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/10321573
• 关键词: 3D Print; Address; Antibiotics; Bacillus; Bacteria; Binding; Biological Assay; Brazil; Case Study; Catalytic DNA; Cessation of life; Child; Chronic Disease; Clinical; Collaborations; Color; Communicable Diseases; Complementarity Determining Regions; 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; Mycobacterium tuberculosis complex; 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; Tuberculosis diagnosis; Validation; bacterial lysate; base; clinically relevant; co-infection; cohort; comorbidity; cost; detection limit; diagnostic assay; diagnostic platform; diagnostic tool; diagnostic value; effective therapy; flexibility; fluorophore; global health; improved; innovation; instrument; isothermal amplification; mortality; mycobacterial; non-tuberculosis mycobacteria; novel; opportunistic pathogen; pathogen; point of care; point of care testing; point-of-care detection; 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.

项目摘要 结核病（TB）是由结核分枝杆菌复合物（MTC）引起的一种传染病， 仍然是全球主要的公共卫生问题之一，有约1000万个新病例，约150万例 每年死亡。巴西在2017年报告了约70,000例新案件，仍然是22个高负荷国家之一 该占全球所有结核病案件的80％。非结婚菌（NTM）是一个新兴组 引起TB样肺部感染的相关机会病原体，特别是在潜在的患者中 合并症，例如HIV或囊性纤维化。 NTM感染臭名昭著，对治疗难治性，<50％ 用口服和可注射的抗生素鸡尾酒治疗某些物种目的地一年的治疗方案 毒品。艾滋病毒共同感染促进疾病进展和 使TB和NTM肺部感染的诊断复杂化。估计有350万例结核病案件未被发现 每年，很大程度上是由于缺乏负担得起的，有效的TB/NTM感染诊断。作为 结果，感染患者患上更严重的疾病，并继续传播这些病原体。频率 通过常用的涂片显微镜方法误诊NTM感染作为结核 可以导致临床结果较差的治疗。这是迫切需要迫切需要的护理（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行数据 疾病。该项目的目的是利用DNA纳米技术的最新进步， 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，二进制的特异性 脱氧核酶（Bidz）传感器以及3D打印测定设备的经济性和可移植性。我们将优化 多色多路复用传感器测定法，以特异性检测痰液样品对TB和NTM病原体的特异性检测。一个 廉价的，便携式电池供电的设备用于测定孵育和使用3D打印的荧光检测 技术将进行测试。最后，将在巴西进行此诊断测定的“现实世界”验证 来自HIV+和HIV-COHORTS的样本。如果成功，可以探索这种灵活的技术来设计POC 其他传染病的诊断测定法。这种国际合作的额外结果将是 增强巴西传染病研究能力和基础设施。