Integrated system for Category C infectious disease diagnostics in resource-limi

项目4的总体目标是开发和验证用于在低资源作业环境中检测耐多药结核病的样本应答系统的性能，将创新和经验证的分子签名转化为现场可部署的诊断测试，并在商业生产之前将系统转移到秘鲁利马进行联合临床前测试和评估。我们将利用获得专利的Akonni制造、样品制备、放大、微流控和微阵列技术，通过三个相互关联的具体目标来实现这些目标：1)开发和验证从净化后的痰中自动提取核酸的方法、消耗品和设备。这一目标还包括从大体积的非侵入性临床标本(如尿液)中提纯核酸的基础科学，并支持哈佛项目2和秘鲁的合作者确定是否可以在儿童和临床样本中可靠地检测到结核杆菌的特征，而不是痰。 2)开发和验证低成本、样品制备和扩增微阵列耗材。耗材设计用于在室温下长期存储，并将保持在低资源环境下进行分子诊断所需的完全封闭的扩增工作流程。这一目标还包括将哈佛项目1和3中创新的MDR和XDR-TB签名转化为下一代可现场部署的诊断测试。 3)与秘鲁利马的哈佛合作者共同开发样本输入、答案输出集成系统(AIMS 1和AIMS 2的硬件、软件和耗材)，用于临床前测试。由此产生的样品制备方法、消耗品和仪器通常将扩展到其他A-C类面板测试，以用于低资源环境。

The overarching objectives of Project 4 are to develop and verify the performance of a sample-in answer-out system for detecting MDR-TB in low-resource operating environments, translate innovative and verified molecular signatures into field deployable diagnostic tests, and transfer systems to Lima Peru for joint, preclinical testing and evaluation ahead of commercial production. We will meet these objectives by taking advantage of patented Akonni manufacturing, sample preparation, amplification, microfluidic, and micorarray technologies through three inter-related specific Aims: 1) Develop and verify methods, consumables, and equipment for automated nucleic acid extraction from decontaminated sputum. This Aim also includes fundamental science for purifying nucleic acids from large-volumen, non-invasive clinical specimens such as urine, and supporting Harvard Project 2 and collaborators in Peru to determine whether or not MTB signatures can be reliably detected in children and in clinical samples other than sputum, 2) Develop and verify low-cost, sample preparation and amplification microarray consumables. Consumables are designed for long-term storage at room temperature, and will maintain an entirely closed-amplicon work flow that is required for molecular diagnostics in low-resource settings. This Aim also includes translating new, innovative MDR- and XDR-TB signatures from Harvard Projects 1 and 3 into a next-generation, field-deployable diagnostic test. 3) Develop sample-in, answer-out integrated system (hardware, software, and consumables from Aims 1 and 2) for pre-clinical testing with Harvard collaborators in Lima, Peru. The resulting sample preparation methods, consumables, and instrument will be generally extensible to other Category A-C panel tests for use in low-resource environments.