Rapid Molecular Detection of Tuberculosis without PCR amplification

项目摘要 结核病（TB）是一个主要的全球负担，每年造成150万人死亡。主要是肺结核 影响发展中国家，但多药和广泛耐药菌株的增加是一个挑战。 全球威胁。为了控制和根除这一疾病，必须拥有适当的工具 在有限资源环境和床旁（POC）环境中检测活动性TB， 大多数结核病患者都能看到。然而，可用的检测方法具有较差的性能， 不适合在有限资源或POC环境中进行诊断。最常用的方法 检测结核病的方法是涂片显微镜检查，这是一种100多年前开发的方法， 能检测出50%的病例 在第一阶段，我们开发了一种快速、稳健的样品制备和检测程序， 易于自动化，并且高度敏感和特异性。新检测方法基于单分子 扫描（SMS），一种能够使用简单的方法检测单个分子的新技术， 设备.在这里，我们将开发一个完全自动化的下一代分子测试的基础上， 第一阶段建立SMS的可行性。我们的目标是取代痰涂片显微镜， 在结核病负担高的国家， 发达国家如果成功的话，新的分析方法将比目前的领导者更准确 分子检测（>95%的灵敏度，>99%的特异性），其价格将是有限的， 资源设置（$6/测试，$1，200/仪器）。此外，新的检测方法将满足所有 根据世卫组织的报告，产品中要求的替代涂片的特性 显微镜 第二阶段的总体目标是开发和验证一次性检测盒和全面的 结核病检测的自动化仪器。Aim 1专注于一次性产品的开发 目标2涉及便携式仪器的开发，目标3涉及核查， 最终仪器和测试卡片设计的临床确认。这一评价与 项目期间积累的设计文档将与其他临床 评估以获得CE标志、FDA批准、结核病高负担国家的批准， 世界卫生组织的认可。

Project Summary Tuberculosis (TB) is a major global burden that kills 1.5 million people every year. TB mostly affects developing countries, but the rise of multi-drug and extensively drug resistant strains is a global threat. In order to control and eradicate this disease, it is imperative to have adequate tools to detect active TB in limited-resource settings and point-of-care (POC) settings, which is where most TB patients are seen. However, available detection methods have poor performance and are not suited for diagnosis in limited-resource or POC settings. The most commonly used method to detect TB is smear microscopy, a procedure developed more than 100 years ago that only detects 50% of the cases. In Phase I, we developed a sample preparation and detection procedure that is rapid, robust, easy to automate, and highly sensitive and specific. The new assay is based on Single Molecule Scanning (SMS), a novel technology capable of detecting single molecules using simple equipment. Here, we will develop a fully automated next generation molecular test based on the feasibility of SMS stablished in Phase I. The goal is to replace smear-microscopy for sputum- based diagnosis of TB in high TB-burden countries and to replace PCR-based TB diagnosis in developed countries. If successful, the new assay will be more accurate than the current leader molecular assay (>95% sensitivity, >99% specificity) and its price will be affordable for limited- resource settings ($6/test, $1,200/instrument). In addition, the new assay will satisfy all the characteristics that according to a WHO report are required in a product to replace smear microscopy. The overall goal of Phase II, is to develop and to validate a disposable cartridge and a fully automated instrument for TB testing. Aim 1 focuses on the development of the disposable cartridge, Aim 2 on the development of the portable instrument and Aim 3 on the verification and clinical validation of the final instrument and cartridge design. This evaluation together with the design documentation accumulated during the project will be combined with additional clinical evaluations to obtain CE mark, FDA approval, approval in high TB-burden countries, and the endorsement of the World Health Organization.