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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）：细菌性脑膜炎是一种严重的细菌感染，可在症状出现后24小时内致命。据世界卫生组织估计，每年发生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昂贵且耗时（例如，细胞培养可能需要48小时以上），要么需要昂贵的专用设备（例如，实时PCR所需的热循环仪）。所有这些因素都限制了这些方法在资源有限的情况下的应用。考虑到脑膜炎多发生在农村高贫困地区，且脑膜炎致死率高，迫切需要一种简单、低成本、高灵敏度的设备和方法，以便在资源贫乏地区对脑膜炎进行即时和早期诊断。该提案的目标是开发一种低成本的医疗点（POC）设备，用于快速和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，特别是在资源贫乏的环境中。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不同DNA序列的DNA编码嫁接到纸上。微流控装置上的多个捕获区允许同时和特异性地检测芯片上这三种不同种类的细菌，基于与特定DNA捕获探针的DNA杂交。将设计一种微型薄膜加热器，将环介导的等温放大（LAMP）集成在芯片上，提供高检测灵敏度。此外，我们还旨在设计和完善一种无仪器的、基于纸张的多发性脑膜炎现场诊断装置。该项目将提供一种低成本的纸质微流体POC设备和方法，用于脑膜炎的即时和早期诊断，特别是在资源贫乏的环境中。它在检测各种植物、动物、食源性疾病和其他微生物的传染病方面也有很大的潜力。