Grafting DNA Codes on Paper for Rapid, Low-Cost, and Sensitive Meningitis Diagnos

将 DNA 代码移植到纸上以实现快速、低成本和灵敏的脑膜炎诊断

基本信息

批准号：
8792227
负责人：
XiuJun Li
金额：
$ 14.79万
依托单位：
UNIVERSITY OF TEXAS EL PASO
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-05-01 至 2017-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8792227
关键词：
Animals Bacteria Bacterial Infections Bacterial Meningitis Bacterial Typing Bedside Testings Biological Brain Cell Culture Techniques Clinical Code Communicable Diseases Complex Consumption DNA DNA Primers DNA Sequence DNA amplification Detection Devices Diagnosis Diagnostic Diphosphates Dyes Early Diagnosis Epidemic Equipment Film Forensic Medicine Genetic Fingerprintings Glass Goals Gram&apos s stain Haemophilus influenzae Health Healthcare Hour Laboratories Life Liquid substance Mediating Membrane Meninges Meningitis Methods Mexico Microfluidic Microchips Microfluidics Neisseria meningitidis Oxides Paper Patient Care Plants Polymerase Chain Reaction Poverty Areas Process Reagent Resources Rural SYBR Green I Sampling Single-Stranded DNA Specificity Spinal Cord Streptococcus pneumoniae Symptoms System Techniques Technology Testing Texas Time World Health Organization accurate diagnosis base biochip cost design foodborne instrument killings microorganism miniaturize nanomaterials pathogen point of care routine Bacterial stain

项目摘要

DESCRIPTION (provided by applicant): Bacterial meningitis is a severe bacterial infection of the protective brain membranes and can become fatal in as early as 24 hours after symptoms are noticed. According to the World Health Organization, one million cases of bacterial meningitis are estimated to occur and 200,000 of these die annually. However, current approaches for meningitis diagnosis are either costly and time consuming (e.g. cell culture, which may take more than 48 hours.), or need expensive specialized equipment available only in laboratories (e.g. thermal cyclers needed for real-time PCR). All these factors limit the applications of those approaches in resource-limited settings. Considering that many cases of meningitis cases happened in rural high-poverty areas, and the high fatality rate of meningitis, a simple, low-cost, highly-sensitive device and method are greatly needed for immediate and early diagnosis of meningitis in resource-poor settings. The goal of this proposal is to develop a low-cost point of care (POC) device for rapid and highly-sensitive diagnosis of three different types of bacterial meningitis, namely Neisseria meningitidis, Haemophilus influenzae, and Streptococcus pneumonia especially for resource-poor settings. DNA codes with different DNA sequences to target these pathogens will be grafted to paper by utilizing the interaction between the nanomaterial graphene oxide (GO) and single strand DNA. Multiple capture zones on microfluidic devices allow for the detection of these three different species of bacteria on chip simultaneously and specifically, based on DNA hybridization with specific DNA capture probes. A miniaturized film heater will be devised to integrate the loop-mediated isothermal amplification (LAMP) on chip, providing high detection sensitivity. In addition, we also aim to design and perfect an instrument-free, paper-based device for multiplex meningitis diagnosis in field. This project will provide a low-cost paper-based microfluidic POC device and method for immediate and early diagnosis of meningitis especially in resource-poor settings. It will also have great potential in the detection of various plant, animal, foodborne, and infectious diseases from other microorganisms.

描述（由申请人提供）：细菌性脑膜炎是保护性脑膜的严重细菌感染，最早在发现症状后 24 小时内即可致命。据世界卫生组织称，估计每年会发生 100 万例细菌性脑膜炎，其中 20 万人死亡。然而，目前的脑膜炎诊断方法要么成本高昂且耗时（例如细胞培养，可能需要超过 48 小时），要么需要实验室才有的昂贵的专用设备（例如实时 PCR 所需的热循环仪）。所有这些因素限制了这些方法在资源有限的环境中的应用。考虑到农村高贫困地区脑膜炎病例较多，且脑膜炎病死率较高，急需一种简单、低成本、高灵敏度的设备和方法，以对资源贫乏地区的脑膜炎进行即时、早期诊断。设置。该提案的目标是开发一种低成本护理点 (POC) 设备，用于快速、高灵敏度地诊断三种不同类型的细菌性脑膜炎，即脑膜炎奈瑟菌、流感嗜血杆菌和肺炎链球菌，特别适用于资源匮乏的环境。利用纳米材料氧化石墨烯 (GO) 和单链 DNA 之间的相互作用，将针对这些病原体的具有不同 DNA 序列的 DNA 代码移植到纸张上。微流体装置上的多个捕获区域允许基于与特定 DNA 捕获探针的 DNA 杂交同时且特异性地检测芯片上的这三种不同细菌。将设计一种小型化薄膜加热器，将环介导等温扩增（LAMP）集成在芯片上，提供高检测灵敏度。此外，我们还致力于设计和完善一种无仪器、纸质的多发性脑膜炎现场诊断设备。该项目将提供一种低成本纸基微流控 POC 设备和方法，用于脑膜炎的即时和早期诊断，特别是在资源匮乏的环境中。它还在检测其他微生物引起的各种植物、动物、食源性疾病和传染病方面具有巨大潜力。