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）：SBIR I期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，用于低资源环境（LRS）的有效分子诊断。这项技术还将有助于监测艾滋病、疟疾和结核病等全球性疾病患者的治疗反应。由于可靠和可重复的高质量NA样品制备过程对于大多数分子诊断分析的成功以及随后的治疗和恢复进展的监测至关重要，因此，如果在非传统或资源匮乏的卫生保健环境中进行测试，则可获得有效的样品制备技术尤为重要。目前商业样品制备方法中的大多数过程都是劳动密集型的，需要特殊的仪器。虽然市售的自动化系统可以进行广泛的样品处理，但在非中心实验室环境中使用它们是昂贵的。一些研究工作集中在“样品输入，答案输出”的微流体全分析系统（¿TAS）的开发上。虽然这些努力通常会产生具有令人印象深刻的分析能力的¿TAS平台，并代表着一种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限制，因为它们需要对LRS来说过于复杂和昂贵的仪器。为此，我们的合作者最近报道了一种便携式，无离心机，室温核酸（NA）提取系统。该系统用于核酸制备（或SNAP）只需要热稳定的试剂，因此该系统与LRS中的存储和运输兼容。与其他需要立即洗脱捕获NA的样品制备技术不同，样品稳定在可拆卸的墨盒上，可以在护理点洗脱，也可以运送到集中实验室进行后期分析。AI Biosciences, Inc. （AIBI）建议使用正在申请专利的多腔室预填充注射器技术，显着减小SNAP技术的尺寸和重量，并减少执行提取所需的总动手时间。这只预充注射器将包含通过SNAP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SPE柱上进行萃取。在第一阶段，我们将演示从实际样品中有效提取，并在15分钟或更短的时间内提供PCR和RT-PCR准备好的模板。分离NA的产量和纯度预计将与那些需要更多仪器和动手时间的市售试剂盒相匹配。我们还将评估在LRS中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取得成功，我们将在第二阶段寻求开发一种低成本、全自动和多样品的系统。