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

基本信息

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
项目类别：
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

项目摘要

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.

抽象的腹泻病是世界范围内的一个主要健康问题，每年导致近 50 万人死亡 5 岁以下的儿童（主要是发展中国家）的存活率更高。生长发育不良和认知发育迟缓的风险胃肠道 (GI) 的识别。目前，腹泻引起疾病的病原体有助于指导有效的治疗。发达国家，这是通过多重 PCR 实验室测试来完成的，这种测试昂贵且缓慢不幸的是，这种实验室测试无法在远程和实验室进行。世界上资源有限的发展中国家地区，腹泻死亡率最高。因此，我们建议开发一种集成的、低成本的即时护理 (POC) 测试来识别 GI 该测试对于资源有限的环境很有用。发展中国家以及发达国家的初级保健机构对于在临床医生办公室实时检测胃肠道病原体非常有用，这样患者就可以立即使用正确的药物进行治疗，而不丢失患者的随访是主要问题。检测胃肠道 (GI) 病原体的一个重要因素是粪便样本的复杂性，通常充满了颗粒物和胆汁盐等分子会抑制信号放大。在之前的 SBIR 项目中，GoDx 已经进行了检测。开发了样品制备化学、DNA 扩增和横向检测分析我们的下一步是开发一种病原体检测方法。完全集成样品处理和信号放大的装置，可用于为了实现这一目标，我们建议开发一种粪便样本制备方法。设备能够可靠地提供用于扩增和检测的可行 DNA，同时需要此处提出的技术将能够提取和丰富内容。只需几分钟即可从粪便拭子中提取 DNA，无需离心机或泵。该项目的第二阶段，我们将全面整合 GoDx 的扩增和检测分析使用此处开发的样品制备技术，提供无缝、最少步骤的 POC 用于检测引起世界各地腹泻病的胃肠道病原体的设备。