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.

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岁以下儿童中。那些幸存下来的人有更高的发育迟缓的风险， 认知发展导致腹泻的胃肠道（GI）病原体的鉴定 疾病有助于指导有效的治疗。目前，在发达国家， 多重PCR实验室测试，其昂贵、缓慢并且需要复杂的高灵敏度。 复杂性实验室不幸的是，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和紧急护理环境 其中在临床医生的办公室中真实的检测GI病原体将是有用的， 病人可以用正确的药物治疗，马上，没有病人的损失，以跟进。的 检测胃肠道（GI）病原体的主要问题是粪便样品的复杂性， 通常充满颗粒和分子如胆汁盐。这些污染物可以抑制 信号放大和检测，如果没有适当地从样品中去除。在以前的SBIR中 GoDx已经开发了样品制备化学，DNA扩增和检测 一组胃肠道病原体的横向流动试纸条测定。我们的下一步是开发一个 病原体检测设备，完全集成了样品处理和信号放大， 其可以在护理点（POC）快速使用。为此，我们建议制订一套 能够稳健地提供用于扩增和检测活DNA的粪便样品制备装置均 同时需要最少的用户动作来这样做。这里提出的技术将能够 在几分钟内从粪便拭子中提取和富集DNA，而无需 离心机或泵。在该项目的第二阶段，我们将完全整合GoDx的放大功能， 和检测分析与样品制备技术在这里开发的，提供了一个无缝， 用于检测引起消化道疾病的GI病原体的最小步骤POC装置 世界各地