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

描述(申请人提供)：细菌性脑膜炎是一种严重的细菌感染的保护性脑膜，最早可在发现症状后24小时内死亡。根据世界卫生组织的数据，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)，要么需要昂贵的专门设备(例如，实时聚合酶链式反应所需的热循环仪)。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。考虑到脑膜炎病例多发生在农村高度贫困地区，且脑膜炎病死率高，迫切需要一种简单、低成本、高灵敏度的设备和方法来在资源匮乏的环境下对脑膜炎进行即时和早期诊断。这项提议的目标是开发一种低成本的护理点(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，特别是在资源匮乏的环境中。利用纳米材料氧化石墨烯(GO)和单链DNA之间的相互作用，具有不同DNA序列的DNA代码将被嫁接到纸上。微流控设备上的多个捕获区允许同时检测芯片上这三种不同的细菌，具体而言，基于与特定DNA捕获探针的DNA杂交。将设计一种小型化的薄膜加热器，将环路介导的等温放大(LAMP)集成在芯片上，提供高检测灵敏度。此外，我们还致力于设计和完善一种现场多路脑膜炎诊断的无仪器、纸质设备。该项目将提供一种低成本的纸基微流控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。它还将在检测来自其他微生物的各种植物、动物、食源性和传染病方面具有巨大的潜力。

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.