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

结核分枝杆菌(MTB)感染了世界三分之一的人口，导致900万人患上结核病 每年有170万人死亡，每年造成170万人死亡。虽然有效的治疗方法 结核病已经存在了50多年，控制它的一个主要障碍仍然是无法 在低资源结核病流行环境中可靠地诊断疾病。大多数临床测试都需要 患者可能很难获得的足够的痰(来自肺部的粘液)样本 没有咳嗽，在儿童和艾滋病毒感染者中。尿液提供了一种诱人的治疗方法 护理点结核病诊断，因为它丰富，收集简单，而且收集不涉及 暴露在传染性气雾剂中。迫切需要减少对结核病的现场诊断 疾病传播和改善治疗结果。一种基于聚合酶链式反应的检测方法的建立 来自尿液的结核病，以及它作为一种护理点检测的发展，可能会对结核病产生巨大的影响 诊断。 目的1.证明从患者尿液中诊断肺结核的能力。我们会 尿液标本检测诊断肺结核的效果评价 七天的治疗开始。我们将计算已知的30个对象的敏感度 根据痰分析结果，计算50例健康对照的特异度。 目标2.通过将目标扩大到结核病基因组的更多区域来提高绩效。 缺乏对结核病基因组中诊断靶点的敏感性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。 我们将结合其他已报告的目标来评估共同目标，以确定一套强有力的目标 为了更高的精度。随着这些检测方法的发展，我们将重新检测AIM 1的尿样。 目的3.演示一种基于定量工程的护理点兼容设备 捕捉化学物质和条件。我们将对业绩进行量化，以允许知情人士 演示原理证明装置的工程学。选择的演示将不会是 最终设备，但将展示与设计目标一致的原则证明：快速 加工，自备货架稳定试剂，无需实验室设备，低成本一次性使用 成分简单，使用步骤简单，从~1毫升尿液中目标回收率为80%。

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.