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操作环境中检测耐多药结核病的样本输入回答系统的性能，将创新和验证的分子特征转化为现场可部署的诊断测试，并将系统转移到利马。秘鲁在商业生产之前进行联合临床前测试和评估。我们将通过三个相互关联的具体目标，利用Akonni专利的制造、样品制备、扩增、微流体和微阵列技术来实现这些目标：1）开发和验证从净化痰液中自动提取核酸的方法、耗材和设备。该目标还包括用于从大体积、非侵入性临床样本（如尿液）中纯化核酸的基础科学，并支持哈佛项目2和秘鲁的合作者，以确定是否可以在儿童和除痰液以外的临床样本中可靠地检测MTB特征， 2)开发和验证低成本的样品制备和扩增微阵列耗材。耗材设计用于在室温下长期储存，并将保持在低资源环境中进行分子诊断所需的完全封闭的扩增子工作流程。该目标还包括将来自哈佛项目1和3的新的、创新的耐多药和广泛耐药结核特征转化为下一代、可现场部署的诊断测试。 3)与秘鲁利马的哈佛合作者一起开发用于临床前测试的样本输入、应答输出集成系统（目标1和2的硬件、软件和耗材）。由此产生的样品制备方法、消耗品和仪器通常可扩展到其他A-C类样本组检测，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.