Point-of-care diagnosis of pulmonary TB from urine samples

通过尿液样本即时诊断肺结核

• 批准号: 9166566
• 负责人: Barry Ryan Lutz
• 金额: $ 27.04万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-03 至 2018-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9166566
• 关键词: Address; Aerosols; Bacillus (bacterium); Base Pairing; Bedside Testings; Biological Assay; Blood; Cells; Cessation of life; Chemistry; Child; Clinical; Clinical Sensitivity; Collection; Complex; Coughing; DNA; Detection; Development; Devices; Diagnosis; Diagnostic; Disease; Engineering; Experimental Designs; Exposure to; Foundations; Genbank; Gene Targeting; Genes; Genome; Genomics; Goals; Gold; Growth; HIV; HIV/TB; Health Status; Healthcare; Immune; Individual; Kidney; Length; Literature; Low income; Lung; Methods; Microscopy; Mucous body substance; Mycobacterium tuberculosis; Nucleic Acids; Patients; Performance; Population; Preparation; Process; Pulmonary Tuberculosis; Reaction; Reagent; Recovery; Reporting; Research Design; Resources; Sampling; Sensitivity and Specificity; Sodium Chloride; Specificity; Sputum; Stream; Testing; Time; Treatment outcome; Tuberculosis; Urine; Work; base; burden of illness; cost; design; disease diagnosis; disease transmission; effective therapy; improved; laboratory equipment; laboratory facility; low and middle-income countries; point of care; treatment duration

M. tuberculosis (MTb) infects one-third of the world’s population, causes TB disease in 9 million people per year, and is responsible for 1.7 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. Point-of-care TB diagnostics are urgently needed to decrease disease transmission and improve treatment outcomes. 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. Demonstrate the ability to diagnose pulmonary TB from patient urine. We will evaluate the performance of detection for diagnosing pulmonary TB from urine of patients within seven days of treatment initiation. We will calculate sensitivity in the 30 subjects known to have pulmonary TB based on sputum analysis results, and calculate specificity in 50 healthy controls. Aim 2. Enhance performance by expanding targets to additional regions of the TB genome. Extensive analysis of the sensitivity and specificity of diagnostic targets in TB genomes is lacking. We will analyze TB genomes in the NCBI GenBank to determine inclusiveness and specificity. We will assess common targets in combination with other reported targets to identify a strong set for high accuracy. As these assays are developed, we will re-test urine samples from Aim 1. Aim 3. Demonstrate a point-of-care-compatible device based on quantitative engineering of capture chemistry and conditions. We will quantify performance to allow informed engineering to demonstrate a proof-of-principle device. The chosen demonstration will not be a final device but will demonstrate a proof-of-principle that is aligned with the design goals: rapid processing, self-contained shelf-stable reagents, no lab equipment, low-cost disposable components, simple user steps, target recovery >80% from ~1 mL urine.

结核分枝杆菌（MTB）感染了世界人口的三分之一，导致900万人结核病疾病 人每年，每年造成170万人死亡。同时有效治疗 结核病已有50多年了，其控制的主要障碍仍然无法 可靠地诊断在低资源结核病环境中。大多数临床测试都需要 足够的痰液样本（肺部粘液）可能难以获得患者 没有咳嗽，儿童和感染了艾滋病毒的人。尿液提供了一种有吸引力的方法 护理点结核病诊断很丰富，收集易于收集，收集不涉及 暴露于感染性气溶胶。迫切需要急需降低的护理结核病诊断 疾病传播和改善治疗结果。开发基于PCR的测试 尿液中的结核病及其作为保健测试的发展可能会对结核病产生巨大影响 诊断。 AIM 1。证明可以从患者尿液中诊断肺结核的能力。我们将 评估从患者尿液中诊断肺结核的检测性能 7天的治疗计划。我们将计算30名已知具有的敏感性 基于痰分析的肺结核结果，并计算50个健康对照中的特异性。 AIM 2。通过将目标扩展到结核病基因组的其他区域来提高性能。 缺乏对结核病基因组诊断靶标的灵敏度和特异性的广泛分析。 我们将分析NCBI GenBank中的结核病基因组，以确定包容性和特异性。 我们将评估共同的目标与其他报告的目标结合使用以确定强大的集合 高准确性。随着这些测定的开发，我们将从AIM 1重新测试尿液样本。 AIM 3。根据定量工程演示一种兼容的兼容设备 捕获化学和条件。我们将量化性能以允许知情 工程学以演示原理设备。选定的示威将不是 最终设备，但将展示与设计目标一致的原理证明：快速 加工，独立的货架稳定试剂，没有实验室设备，低成本处置 组件，简单的用户步骤，目标恢复> 80％从〜1 ml尿液中。