Rapid, Handheld, Point-of-Care Stool Sample-prep Device for the Extraction and Detection of Nucleic Acid from Diarrheal Pathogens

用于从腹泻病原体中提取和检测核酸的快速、手持式护理点粪便样本制备装置

• 批准号: 10086785
• 负责人: Chang Hee Kim
• 金额: $ 29.94万
• 依托单位: GODX, INC.
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-01 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10086785
• 关键词: 3D Print; 5 year old; Activated Charcoal; Age; Bedside Testings; Binding; Biological Assay; Buffers; Bulla; Centrifugation; Cessation of life; Chemistry; Child; Complex; Coupling; Cytolysis; DNA; DNA Binding; DNA amplification; Detection; Development; Devices; Diagnostic; Diarrhea; Environment; Excision; Feces; Genes; Goals; Growth and Development function; Health; Human Resources; Industry Standard; Ion Exchange Resins; Laboratories; Lateral; Manuals; Methods; Microfluidics; Microprocessor; Microspheres; Molecular; Motor; Nucleic Acids; Paper; Particulate; Pathogen detection; Patients; Pharmaceutical Preparations; Phase; Plant Resins; Porifera; Preparation; Price; Primary Health Care; Process; Protocols documentation; Pump; Reagent; Recovery; Reporting; Resources; Sampling; Signal Transduction; Silicon Dioxide; Small Business Innovation Research Grant; Solid; Swab; System; Techniques; Technology; Testing; Time; Training; Work; amplification detection; base; bile salts; cognitive development; cost; design; diarrheal disease; effective therapy; follow-up; gastrointestinal; high risk; inhibitor/antagonist; instrument; laboratory equipment; mortality; multiplex detection; novel; nucleic acid detection; pathogen; point of care; primary care setting; stool sample

Abstract Diarrheal disease is a major health issue worldwide resulting in nearly 500,000 deaths annually in children under 5 years of age, mostly in the developing world. Those who survive have higher risk of stunted growth and cognitive development. Identification of the gastrointestinal (GI) pathogens that cause diarrheal disease helps guide effective treatment. Currently, in the developed world, this is done using multiplexed PCR laboratory tests which are expensive, slow and require a laboratory. Unfortunately, such laboratory tests are not available in remote and resource limited regions in the developing world where mortality due to diarrhea is the highest. Thus, we propose to develop an integrated, low-cost, point-of-care (POC) test to identify the GI pathogens that cause diarrheal diseases. This test will be useful in resource-limited settings of the developing world as well as in primary care settings of the developed world where it will be useful to detect the GI pathogens in real time, in the clinician's office so the patient can be treated with the right drugs, right away, without loss of patients to follow up. The major problem with detecting gastrointestinal (GI) pathogens is the complexity of stool samples, often filled with particulate and molecules such as bile salts. Such contaminants can inhibit signal amplification and detection if not properly removed from the sample. In previous SBIR projects, GoDx has developed sample preparation chemistry, DNA amplification and detection assays on lateral flow paper strips for a suite of GI pathogens. Our next step is to develop a pathogen detection device that fully integrates sample processing and signal amplification, which can be used rapidly at the point of care (POC). To achieve this, we propose to develop a stool sample prep device that can robustly provide viable DNA for amplification and detection all while requiring minimal user action to do so. The technology proposed here will be able to extract and enrich DNA from a swab of stool in a matter of minutes without the need for centrifuges or pumps. In Phase 2 of this project, we will fully integrate the GoDx’s amplification and detection assays with the sample prep technology developed here, providing a seamless, minimal step POC device for the detection of GI pathogens that cause diarrheal diseases around the world.

摘要 腹泻病是世界范围内的主要健康问题，每年导致近50万人死亡 在5岁以下的儿童中，主要是在发展中国家。那些幸存下来的人有更高的 发育迟缓和认知发展的风险。胃肠道（GI）的识别 病原体引起的疟疾疾病有助于指导有效的治疗。目前在 在发达国家，这是使用多重PCR实验室测试来完成的，这是昂贵的，缓慢的， 需要一个实验室不幸的是，这种实验室测试在远程和远程环境中不可用。 在发展中国家的资源有限地区，腹泻造成的死亡率最高。 因此，我们建议开发一种集成的、低成本的床旁（POC）检测来识别GI 导致尿道疾病的病原体。这个测试将是有用的资源有限的设置， 发展中国家以及发达国家的初级保健环境， 用于在临床医生的办公室中真实的实时检测GI病原体， 用正确的药物治疗，马上，没有病人的损失，以跟进。的主要问题 与检测胃肠道（GI）病原体有关的是粪便样本的复杂性，通常充满了 微粒和分子如胆汁盐。这些污染物会抑制信号放大 以及如果没有从样品中适当地去除则检测。在以前的SBIR项目中，GoDx 开发了样品制备化学、DNA扩增和检测分析， 一系列胃肠道病原体的流动试纸我们的下一步是开发一种病原体检测 完全集成样品处理和信号放大的设备，可用于 在护理点（POC）快速。为了实现这一目标，我们建议开发一种粪便样本制备方法， 一种可以稳健地提供用于扩增和检测的可行DNA的装置， 最少的用户操作来完成此操作。这里提出的技术将能够提取和浓缩 在几分钟内从粪便拭子中提取DNA，而无需离心机或泵。在 在该项目的第二阶段，我们将完全整合GoDx的扩增和检测分析， 利用这里开发的样品制备技术，提供无缝、最小步骤的POC 用于检测全球范围内引起肠道疾病的GI病原体的设备。