A Multiplex Nanodiagnostic Array for Detection of Emerging Pathogens

描述（由申请方提供）：新出现的传染病以及故意使用病原体或其毒素进行恐怖行为，构成了全球社会面临的一些最重大的健康和安全挑战。一个关键的挑战是制定有效的战略，以迅速发现和控制自然发生的传染病爆发。特别是对于生物防御和风险评估，快速和明确地识别病原体及其毒素具有巨大的意义。实时、多路复用能力在用于监测、确认和及时实施预防性和保护性公共卫生措施的任何诊断中是必不可少的;然而，常规诊断无法以允许在有限资源、护理点设置中部署的格式提供这些能力。因此，迫切需要稳健的多重策略来鉴定病原体和毒素，其可以容易地部署在几乎任何环境和临床环境中并且容易配置（例如，针对地理病原体血清群和其预期针对的人群）。为了解决这些问题，该项目建议制造和建立下一代原型概念验证，用于检测选择的生物防御和新兴病原体的多重诊断。创新的传感器设计将构成一个高度有序的离散阵列，电寻址纳米同轴传感器阵列，每个具有指定的分子印迹（MI）纳米涂层，用于识别不同的毒素或病原体。这种新型传感器阵列将实现：1）由于其分子尺度和结构，允许空间离散，高站点密度，因此具有多路复用和校对能力（~108/cm 2），电寻址纳米同轴传感器; 2）前所未有的灵敏度和特异性;以及3）提供无标记检测的全电子电容测量，理想地适用于有限的资源，在以前无法实现的规模上进行适合现场的设置。R21组件将制造含有能够超灵敏和特异性毒素识别的MI的纳米同轴传感器，并将建立使用单个多路复用芯片上的纳米同轴传感器阵列同时检测霍乱和滋贺毒素的概念验证。在这些结果的基础上，R33部分将寻求评估通过分子印迹纳米同轴传感器阵列检测病原生物体的概念验证，使用属于O 1和O 139血清群的霍乱弧菌菌株和产滋贺病原体的大肠杆菌（STEC）O 157：H7。R33组件还将设计样品输送和集成电子设备到原型手持式诊断仪中，用于在单个一次性多路芯片上同时检测病原体和毒素。

DESCRIPTION (provided by applicant): Emerging infectious diseases together with the intentional use of pathogenic organisms or their toxins for acts of terror constitute some of the most significant health and security challenges facing the global community. A key challenge is to develop effective strategies to rapidly detect and control naturally occurring outbreaks of infectious disease. Especially for biodefense and risk evaluation, rapid and definitive identification of pathogens and their toxins is of enormous significance. Real-time, multiplexing capabilities are essential in any diagnostic for surveillance, confirmation, and timely implementation of preventive and protective public health measures; however, conventional diagnostics fail to provide these capabilities in a format that allows for deployment in limited resource, point-of-care settings. Consequently, there is a critical need for robust multiplex strategies to identify pathogens and toxins that can be readily deployed in virtually any environmental and clinical setting and easily configured (e.g., targeted for geographic pathogen serogroups and human populations for which it is intended). To address these issues, this project proposes to fabricate and establish proof-of-concept of a prototype next generation, multiplex diagnostic for detection of select biodefense and emerging pathogens. The innovative sensor design will constitute a highly ordered array of discrete, electrically addressable nanocoaxial sensors arrays, each with a specified molecular imprint (MI) nanocoating for recognition of a distinct toxin or pathogen. This novel sensor array will enable: 1) multiplexing and proofreading capability, due to its molecular scale and architecture that allows for spatially discrete, high site density (~108 per cm2), electrically addressable nanocoaxial sensors; 2) unprecedented sensitivity and specificity; and 3) all-electronic capacitance measurement that affords label-free detection, ideally suited for limited resource, field-appropriate settings on a scale that has not previously been achievable. The R21 component will fabricate nanocoaxial sensors containing MIs capable of ultrasensitive and specific toxin recognition and will establish proof-of-concept for the simultaneous detection of cholera and shiga toxins using nanocoaxial sensor arrays on a single multiplex chip. Expanding on these results, the R33 component will seek to evaluate proof-of-concept for detection of pathogenic organisms by molecular imprinted nanocoaxial sensor arrays, using Vibrio cholerae strains belong to the O1 and O139 serogroups and Shiga pathogen-producing Escherichia coli (STEC) O157:H7. The R33 component will also engineer sample delivery and integrated electronics into a prototype handheld diagnostic for simultaneous detection of both pathogens and toxins on a single disposable multiplex chip.