Detection of transrenal Mycobacterium tuberculosis DNA in urine

尿液中结核分枝杆菌DNA的经肾检测

• 批准号: 10261391
• 负责人: Molly Forrest Franke
• 金额: $ 8.45万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-11 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10261391
• 关键词: Address; Aftercare; Antitubercular Agents; Apoptotic; Aspirate substance; Bacteriology; Blood Circulation; Cells; Cessation of life; Characteristics; Child; Clinical; Cohort Studies; DNA; Data; Databases; Deglutition; Detection; Diabetes Mellitus; Diagnosis; Diagnostic; Diagnostic tests; Disease; Feces; Filtration; HIV; Health; Individual; Inpatients; Integration Host Factors; Kidney; Knowledge; Laboratories; Lateral; Measures; Microscopy; Molecular; Monitor; Morbidity - disease rate; Mycobacterium tuberculosis; Mycobacterium tuberculosis antigens; Nucleic Acids; Oral; Patients; Persons; Peru; Population; Recording of previous events; Sampling; Specificity; Sputum; Swab; Symptoms; Testing; Thoracic Radiography; Time; Tuberculin Test; Tuberculosis; Urinalysis; Urine; Work; accurate diagnostics; base; cell free DNA; comorbidity; global health; infection risk; mortality; study characteristics; tool; treatment effect; treatment response; tuberculosis diagnostics; tuberculosis treatment

Project Summary Tuberculosis (TB) is a major global health burden and led to the death of 1.6 million people in 2017. A major driver of this high mortality rate is delay in timely diagnosis. Conventional TB diagnostic tests such as smear microscopy or culture used to confirm TB disease rely on sputum, but many TB patient have culture-negative disease or cannot produce sputum (e.g., children or people living with HIV), which hampers case detection and treatment initiation. Therefore, there is an urgent need for diagnostic tools using other sample types to confirm TB disease. Fragments of cell-free Mycobacterium tuberculosis (Mtb) DNA can pass the kidneys and be detected in urine as transrenal (tr) DNA, offering promise as a diagnostic tool. However, test sensitivity has been variable and factors influencing presence of bacterial trDNA are unknown. As part of a large cohort study in Lima, Peru, we have collected urine samples from TB patients and extracted DNA from these samples. Our preliminary data suggest that levels of Mtb trDNA increase shortly after initiation of treatment due to increased bacterial killing leading to release of nucleic acids in the bloodstream. In this study, we will (1) conduct short-fragment real-time PCR to detect changes in Mtb trDNA over time in extracts from 200 TB cases, of whom we have three serial samples, collected pre-, four, and seven days post treatment start; and (2) associate levels of Mtb trDNA in urine with host factors (such as smear status or comorbidities) and urine factors. This work will provide important information to develop strategies for trDNA-based tools to confirm TB disease and monitor TB treatment response.

项目摘要 结核病（TB）是全球重大的健康负担，导致160万人死亡 2017年。这种高死亡率的主要驱动力是及时诊断的延迟。常规结核病 诊断测试（例如用于确认结核病疾病的涂片显微镜或培养物）依赖 痰液，但许多结核病患者患有培养性疾病或无法产生痰液（例如， 儿童或艾滋病毒感染者），这会阻碍病例检测和治疗启动。 因此，迫切需要使用其他样本类型确认结核的诊断工具 疾病。 无细胞分枝杆菌结核（MTB）DNA的碎片可以通过肾脏并BE 在尿液中检测为跨肾（TR）DNA，提供了作为诊断工具的承诺。但是，测试 敏感性是可变的，影响细菌trDNA存在的因素尚不清楚。 作为秘鲁利马市的一项大型队列研究的一部分，我们收集了结核病患者的尿液样本 并从这些样品中提取DNA。我们的初步数据表明MTB trDNA的水平 因细菌杀死增加而开始治疗后不久增加 血液中的核酸。在这项研究中，我们将（1）实时进行短片段 PCR在200 TB病例的提取物中检测MTB trDNA随时间的变化，我们 三个连续样品，在治疗后收集了前，四个和七天的开始； （2）合伙人 尿液中MTB TrDNA的水平，具有宿主因子（例如涂片状态或合并症）和尿液 因素。这项工作将提供重要的信息，以制定基于trdna的工具的策略 确认结核病并监测结核病治疗反应。