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

• 批准号: 8475160
• 负责人: XiuJun Li
• 金额: $ 14.47万
• 依托单位: UNIVERSITY OF TEXAS EL PASO
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-05-01 至 2016-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8475160
• 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 Fingerprinting; DNA Primers; DNA Sequence; DNA amplification; Detection; Devices; Diagnosis; Diagnostic; Diphosphates; Dyes; Early Diagnosis; Epidemic; Equipment; Film; Forensic Medicine; Glass; Goals; Gram' s stain; Haemophilus influenzae; 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; base; biochip; cost; design; foodborne; instrument; killings; microorganism; miniaturize; nanomaterials; pathogen; point of care; public health relevance; routine Bacterial stain

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保护性脑膜细菌感染，在发现症状后24小时内即可致命。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，目前用于脑膜炎诊断的方法要么昂贵且耗时（例如，细胞培养，其可能需要超过48小时），或者需要仅在实验室中可用的昂贵的专用设备（例如实时PCR所需的热循环仪）。所有这些因素都限制了这些方法在资源有限的环境中的应用。鉴于脑膜炎病例多发生在农村高度贫困地区，且病死率高，迫切需要一种简单、低成本、高灵敏度的设备和方法，用于资源匮乏地区脑膜炎的即时和早期诊断。该提案的目标是开发一种低成本的床旁（POC）设备，用于快速和高灵敏度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，特别是在资源匮乏的环境中。通过利用纳米材料氧化石墨烯（GO）和单链DNA之间的相互作用，将具有不同DNA序列的DNA代码移植到纸上，以靶向这些病原体。微流体装置上的多个捕获区允许基于与特异性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种类的细菌。设计了一种小型化的薄膜加热器，将环介导等温扩增（LAMP）集成在芯片上，提供高的检测灵敏度。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装置。该项目将提供一种低成本的纸基微流体POC装置和方法，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。它也将在检测各种植物，动物，食源性和其他微生物引起的传染病方面具有巨大的潜力。