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: 10611784
• 负责人: Chang Hee Kim
• 金额: $ 105万
• 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 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10611784
• 关键词: 5 year old; Bacteria; Biological Assay; Campylobacter; Cause of Death; Cell Culture Techniques; Centrifugation; Cessation of life; Chemistry; Child; Clinical; Clinical Research; Complex; Contractor; DNA; DNA amplification; Detection; Development; Devices; Diagnostic; Diarrhea; Docking; Dysentery; Electronics; Elements; Engineering; Enteral; Environment; Exodeoxyribonuclease III; Feces; Freeze Drying; Future; Gastroenteritis; Growth and Development function; Health; Heating; Industry Standard; Intake; Laboratories; Lateral; Legal patent; Life; Manuals; Mechanics; Methods; Motor; Nucleic Acids; Outcome; Output; Paper; Particulate; Pathogen detection; Patients; Pharmaceutical Preparations; Phase; Plant Resins; Preparation; Price; Primary Care; Process; Pump; Resource-limited setting; Salmonella; Sampling; Sensitivity and Specificity; Shigella; Signal Transduction; Small Business Innovation Research Grant; Solid; Swab; Technology; Temperature; Testing; Time; Validation; Visual; World Health Organization; amplification detection; bile salts; cognitive development; commercialization; cost; design; detection assay; diarrheal disease; effective therapy; efficacy validation; follow-up; gastrointestinal; high risk; improved; inhibitor; instrument; isothermal amplification; lateral flow assay; manufacture; microdevice; mortality; nucleic acid detection; pathogen; pathogenic bacteria; point of care; point of care testing; stool sample; urgent care

Abstract Diarrheal disease is a major health issue worldwide resulting in nearly 500,000 deaths annually in children under 5 years of age. 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 sophisticated high complexity 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 and urgent 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.

抽象的 腹泻病是全球一个主要的健康问题，每年导致近500,000人死亡 在5岁以下的儿童中。那些生存的人的增长风险更高， 认知发展。引起腹泻的胃肠道（GI）病原体的鉴定 疾病有助于指导有效的治疗。目前，在发达世界中，这是使用 多路复用的PCR实验室测试，昂贵，缓慢并且需要高级高 复杂性实验室。不幸的是，此类实验室测试在遥控器中不可用， 在发展中国家的资源有限的地区，由于腹泻导致的死亡率最高。 这，我们建议开发综合，低成本的护理（POC）测试以识别GI 引起腹泻疾病的病原体。该测试将在资源有限的设置中有用 发展中国家以及发达国家的初级保健和紧急护理环境 在临床上的办公室实时检测胃肠道病原体将很有用 可以立即使用正确的药物治疗患者，而不会失去患者的随访。这 检测胃肠道（GI）病原体的主要问题是粪便样本的复杂性， 通常充满特殊性和分子，例如胆汁盐。这样的污染物可以抑制 信号放大和检测如果未正确从样品中删除。在以前的SBIR中 项目，GODX开发了样品制备化学，DNA扩增和检测 在侧面流纸条上进行一套胃肠道病原体的测定。我们的下一步是开发 病原体检测设备完全整合样品处理和信号放大， 可以在护理点（POC）迅速使用。为了实现这一目标，我们建议开发 凳子样品准备装置，可以强大地提供可行的DNA进行扩增和检测 同时需要最小的用户行动才能做到这一点。这里提出的技术将能够 在几分钟内提取并富含粪便的DNA，而无需 离心机或泵。在该项目的第二阶段中，我们将完全整合GODX的放大 并使用此处开发的样本准备技术进行检测测定，提供了无缝的， 最小的步进POC装置，用于检测引起腹泻疾病的胃肠道病原体 世界各地。