A Portable, Self-Contained Nucleic Acid Extraction and Storage System for POC Tes

用于 POC Tes 的便携式独立核酸提取和存储系统

• 批准号: 8712053
• 负责人: SEASON S-S WONG
• 金额: $ 20.97万
• 依托单位: AI BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-04-01 至 2015-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8712053
• 关键词: Acquired Immunodeficiency Syndrome; Area; Bicycling; Biological Assay; Blood; Cold Chains; Communicable Diseases; Communities; Country; DNA; Detection; Developed Countries; Development; Devices; Diagnosis; Diagnostic; Disease; HIV; HIV Infections; Hand; Healthcare; Injection of therapeutic agent; Laboratories; Legal patent; Location; Malaria; Methods; Microfluidics; Molecular; Monitor; Motor; Nucleic Acids; Patients; Performance; Perinatal; Phase; Point-of-Care Systems; Polymerase Chain Reaction; Preparation; Price; Process; Pump; RNA; Reagent; Recovery; Reporting; Research; Resources; Sampling; Small Business Innovation Research Grant; Solid; Solutions; Spottings; Syringes; System; Systems Analysis; Technology; Temperature; Testing; Time; Tube; Tuberculosis; Viral Load result; Weight; Work; cost; design; disease diagnosis; instrument; instrumentation; point of care; public health relevance; skills; success; treatment response

DESCRIPTION (provided by applicant): This SBIR Phase I proposal aims to develop and commercialize a portable, easy-to-use system for nucleic acid preparation for effective molecular diagnostics in low-resource settings (LRS). The technology will also aid in the monitoring of treatment response for patients of global diseases such as AIDS, malaria, and tuberculosis. Because a reliable and reproducible high-quality sample preparation process for NA is critical to the success of most molecular diagnostic assays and the subsequent treatment and monitoring of recovery progress, the availability of an effective sample preparation technology is especially important if the tests are performed in non-traditional or low-resource health care settings. Most processes in current commercial sample preparation methods are labor intensive and require special instrumentation. Although commercially available automated systems can perform extensive sample processing, they are expensive for use in non-central laboratory settings. Several research efforts have focused on the development of a "sample in, answer out" microfluidic total analysis system (¿TAS). While these efforts have often resulted in ¿TAS platforms with impressive analytical capabilities and represent a potentially paradigm-shifting long term solution, they remain an area of active research and may not be applicable to LRS settings. For instance, the use of these systems is inhibited by their continuing need for instrumentation that is too sophisticated and costly for LRS. To this end, a portable, centrifuge-free, room temperature nucleic acid (NA) extraction system has recently been reported by our collaborator. This system for nucleic acid preparation (or SNAP) requires only thermally stable reagents so the system is compatible with storage and transport in LRS. Unlike other sample preparation technology in which the captured NA needs to be eluted immediately, samples are stabilized on detachable cartridges, which can be eluted either at the point of care, or transported to a centralized laboratory for later analysis. Using a patent pending, multi-chamber, and prefilled syringe technology, AI Biosciences, Inc. (AIBI) proposes to significantly reduce the size and weight of the SNAP technology and to reduce the total hands-on time needed to perform an extraction. This single, pre-filled syringe will contain all the reagents required to conduct DNA/RNA extraction via the solid phase extraction (SPE) column in the SNAP. The actuation of a single plunger can sequentially deliver the necessary reagents to this SPE column to perform an extraction. In Phase I, we will demonstrate effective extraction from realistic samples and to deliver PCR and RT-PCR ready templates in 15 min or less. The yield and purity of the isolated NA are expected to match those obtained from commercially available kits that require far more instruments and hands-on time. We will also assess the long term stability of the pre-filled reagents for practical use in LRS. If the work in Phase I is successful we will seek to develop a low-cost, fully automated, and multi-sample system in Phase II.

描述（由申请人提供）：该 SBIR 第一阶段提案旨在开发和商业化一种便携式、易于使用的核酸制备系统，用于在资源匮乏环境（LRS）中进行有效的分子诊断。该技术还将有助于监测艾滋病、疟疾和结核病等全球疾病患者的治疗反应。由于可靠且可重复的高质量 NA 样品制备过程对于大多数分子诊断测定以及后续治疗和恢复进度监测的成功至关重要，因此，如果测试是在非传统或资源匮乏的医疗机构中进行，则有效的样品制备技术的可用性尤其重要。当前商业样品制备方法中的大多数过程都是劳动密集型的，并且需要特殊的仪器。尽管商用自动化系统可以执行广泛的样品处理，但在非中心实验室环境中使用它们的成本昂贵。多项研究工作的重点是开发“样本输入，答案输出”微流体总体分析系统（¿TAS）。虽然这些努力往往导致 ¿TAS 平台具有令人印象深刻的分析能力，并代表了潜在的范式转变的长期解决方案，但它们仍然是一个活跃的研究领域，可能不适用于 LRS 设置。例如，这些系统的使用受到对仪器仪表的持续需求的限制，而仪器仪表对于远程监视系统而言过于复杂且昂贵。为此，我们的合作者最近报道了一种便携式、免离心、室温核酸（NA）提取系统。该核酸制备（或 SNAP）系统仅需要热稳定试剂，因此该系统与 LRS 中的储存和运输兼容。与其他需要立即洗脱捕获的 NA 的样品制备技术不同，样品稳定在可拆卸的柱上，可以在护理点洗脱，或运输到中央实验室进行后续分析。 AI Biosciences, Inc. (AIBI) 使用正在申请专利的多室预装注射器技术，旨在显着减小 SNAP 技术的尺寸和重量，并减少执行提取所需的总操作时间。这个单一的预填充注射器将包含通过 SNAP 中的固相萃取 (SPE) 柱进行 DNA/RNA 提取所需的所有试剂。单个柱塞的驱动可以顺序地将必要的试剂输送到该 SPE 柱以执行萃取。在第一阶段，我们将展示从真实样品中的有效提取，并在 15 分钟或更短的时间内提供 PCR 和 RT-PCR 就绪模板。分离的 NA 的产率和纯度预计与从需要更多仪器和实践时间的市售试剂盒中获得的产量和纯度相匹配。我们还将评估 LRS 实际使用的预填充试剂的长期稳定性。如果第一阶段的工作成功，我们将在第二阶段寻求开发低成本、全自动、多样本的系统。