Transrenal DNA for point-of-care diagnosis of TB

用于结核病即时诊断的经肾 DNA

• 批准号: 10285973
• 负责人: Barry Ryan Lutz
• 金额: $ 21.14万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-02 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10285973
• 关键词: Adult; Aerosols; Base Pairing; Biological Assay; Cells; Cessation of life; Child; Childhood; Clinical; Clinical Sensitivity; Collection; Communicable Diseases; Coughing; DNA; Detection; Development; Diagnosis; Disease; Exhibits; Exposure to; Foundations; Genomic Segment; Gold; Growth; HIV; HIV Seronegativity; HIV Seropositivity; HIV/TB; Individual; Kidney; Length; Libraries; Low income; Lung; Methods; Microscopy; Mucous body substance; Mycobacterium tuberculosis; Outcome Study; Patients; Population; Preparation; Pulmonary Tuberculosis; Recovery; Resources; Sample Size; Sampling; Sensitivity and Specificity; South Africa; Specificity; Specimen; Sputum; Testing; Time; Tuberculosis; United States; Urine; Work; accurate diagnosis; base; blood filter; co-infection; cohort; design; diagnostic accuracy; disease diagnosis; effective therapy; human DNA; improved; laboratory development; laboratory facility; low and middle-income countries; multiplex assay; multiplex detection; non-tuberculosis mycobacteria; novel; point of care; point of care testing; point-of-care diagnosis; tuberculosis diagnostics; tuberculosis treatment

ABSTRACT M. tuberculosis (Mtb) infects one-third of the world’s population, causes TB disease in 10 million people per year, and is responsible for 1.6 million deaths annually. While effective treatments for TB have been available for over 50 years, a major barrier to its control has remained the inability to reliably diagnose disease in low-resource TB-endemic settings. Most clinical tests require an adequate sample of sputum (mucus from the lungs) which may be difficult to obtain in patients without a cough, in children, and in HIV-infected individuals. Urine offers an attractive method for point-of-care TB diagnostics since it is abundant, simple to collect, and collection does not involve exposure to infectious aerosols. Transrenal DNA (trDNA) present in the urine offers a specific target that can be amplified for high sensitivity. The development of a PCR-based test of TB from urine, and its development as a point-of-care test, could have enormous impact on TB diagnostics. Aim 1. To improve detection of Mtb-specific trDNA using ultrashort PCR and multiplexing. Hypothesis: Amplification of shorter fragments and/or targeting multiple unique fragments will enable detection of low- concentrations samples, increasing sensitivity while maintaining specificity. Aim 2. To determine sensitivity and specificity of improved Mtb-specific trDNA detection in urine samples from adults and children in a TB-endemic setting. Hypothesis: Mtb trDNA is present in detectable quantities in urine from both adults and children with pulmonary TB regardless of HIV status, and our hybridization sequence-capture method can increase clinical sensitivity compared to previous TB trDNA studies.

摘要 M.结核病（Mtb）感染了世界三分之一的人口，每年导致1000万人患结核病， 每年造成160万人死亡虽然结核病的有效治疗方法已经存在了超过20年， 50年来，其控制的一个主要障碍仍然是无法在低资源环境中可靠地诊断疾病。 结核病流行的环境。大多数临床测试需要足够的痰样本（来自肺部的粘液）， 在没有咳嗽的患者、儿童和HIV感染者中可能难以获得。尿液提供了 这是一种有吸引力的护理点TB诊断方法，因为它丰富，易于收集，并且收集不 涉及接触传染性气溶胶。存在于尿液中的跨肾DNA（trDNA）提供了一种特异性靶点， 可以放大以获得高灵敏度。尿结核病PCR检测方法的建立及其进展 作为一种即时检测，可能会对结核病诊断产生巨大影响。 目标1.目的：提高应用超短PCR和多重PCR检测结核分枝杆菌特异性trDNA的效率。假设： 较短片段的扩增和/或靶向多个独特片段将使得能够检测到低表达的DNA。 浓度样品，提高灵敏度，同时保持特异性。 目标二。确定改良的尿结核分枝杆菌特异性trDNA检测的敏感性和特异性 来自结核病流行环境中成人和儿童的样本。假设：Mtb trDNA存在于可检测的 无论艾滋病毒感染状况如何， 与以前TB trDNA研究相比，杂交序列捕获方法可以增加临床灵敏度。