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

M.结核病（MTb）感染了世界三分之一的人口，导致900万人患结核病 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，控制结核病的一个主要障碍仍然是 在资源匮乏的结核病流行环境中可靠地诊断疾病。大多数临床试验需要 可能难以从患者体内获得的足够的痰液样本（来自肺部的粘液） 在儿童和HIV感染者中，尿液提供了一种有吸引力的方法， 因为它丰富，易于收集，并且收集不涉及 接触传染性气溶胶迫切需要护理点结核病诊断， 疾病传播和改善治疗效果。建立了一种基于PCR的 尿液结核病及其作为即时检测的发展可能对结核病产生巨大影响 诊断 目标1。展示从病人尿液中诊断肺结核的能力。我们将 评价检测肺结核病人尿液的性能， 治疗开始7天。我们将计算30个已知有 肺结核的痰液分析结果的基础上，并计算特异性在50名健康对照。 目标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。 对结核病基因组中诊断靶点的敏感性和特异性缺乏广泛的分析。 我们将分析NCBI基因库中的TB基因组，以确定包容性和特异性。 我们将结合其他报告的目标评估共同目标，以确定一套强有力的目标 为了高精度。随着这些检测方法的开发，我们将重新检测目标1的尿液样本。 目标3.展示基于定量工程的床旁兼容设备 捕获化学和条件。我们将量化性能， 工程来演示原理验证设备。选择的演示将不会是 最终器械，但将证明符合设计目标的原理验证：快速 处理，独立储存稳定的试剂，无需实验室设备，低成本一次性使用 组分，简单的用户步骤，目标回收率> 80%，约1 mL尿液。

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.