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.

项目摘要 结核病（TB），这是由结核分枝杆菌复合物（MTC）引起的感染疾病， 仍然是全球主要的公共卫生问题之一，有约1000万个新的卡斯，约150万 每年的死亡人数约有70,000个新案件 该占全球所有结核病的80％。 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 诸如HIV或囊性FIV兄弟之类的合并症。 尽管长达一年的治疗方案和口服和可注射的抗生素鸡尾酒，但物种的治疗率 毒品进一步加剧了这种情况。 将结核病和NTM肺部感染的诊断复杂化。 每年，由于缺乏负担得起的TB/NTM感染的有效的护理诊断 结果，感染的患者患有更严重的疾病，并继续传播这些病原体 通过常用的涂片方法延迟批准，误诊NTM感染作为结核病 可能导致临床结果较差的治疗。 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。 分子传感器和3D打印，可显着提高TB和NTM的POC诊断能力 平台结合了RPA同胸扩增的速度和灵敏度，二进制的特异性 Deoxyribozyme（Bidz）传感器以及3D打印机设备的经济性和便携性 多色多路复用传感器，用于从痰液样品中特异性检测TB和NTM病原体。 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 最终将测试技术的“现实世界”验证该诊断测定法 艾滋病毒+和艾滋病毒的样本。 对其他传染性疾病的诊断测定 增强巴西传染病研究能力和基础设施。