Integrated Microfluidic Platform for Point-of-Care Serodiagnostics

描述(由申请人提供)：我们的目标是开发一个完全集成的微流控免疫诊断平台，利用由Abe Lee博士的实验室首创的新型气液腔声换能器(ALCAT)技术。该诊断平台是基于成熟的临床诊断工具--酶免疫分析(如酶联免疫吸附试验)。ELISA是临床诊断的一个不可或缺的平台，因为它们快速、可靠，并提供高度敏感和特异的结果。此外，ELISA依赖于宿主固有的体液免疫反应来产生指示病理状态的抗体。在这项提议中，我们将结成蛋白质微阵列(Antigen Discovery，Inc.和Phil Felgner博士)和微流控(Abe Lee博士)专业知识的战略合作伙伴关系，以开发一种由成本效益高的一次性微流控塑料芯片和便携式分析仪组成的护理点诊断系统，该分析仪可以由移动计算设备驱动，用于对类梅毒进行快速血清学评估。通过将基于ALCAT的新方法协同整合到这些阵列中，我们将拥有减少酶开发时间、限制生产成本并将所有成分完全集成到一次性检测中的多功能性和能力。从NIH/NIAID以往的资助中，我们已经发现了12种NIAID优先病原体(布鲁氏菌、图拉氏菌、假鼻疽伯克霍尔德氏菌、柯克斯体、伤寒沙门氏菌、立克次体、东方体、弓形虫、结核分枝杆菌、与天花相关的主要痘病毒、登革热和西尼罗河)的血清诊断抗原。我们还鉴定了非生物防御靶标的血清诊断抗原，包括：亨氏巴尔通体、伯氏疏螺旋体、乙肝病毒、衣原体、恶性疟原虫、人乳头瘤病毒和单纯疱疹病毒。这种方法将利用我们现有的血清诊断抗原内容(和适当的银行血清收集)来生产一种多路设备，该设备将包含来自多种病原体的多种抗原，并使临床医生能够确定个人接触了哪种病原体。我们相信，该平台利用了这些丰富的内容，并将其与基于ALCAT的多功能微流体方法相结合，以生产真正的芯片实验室工具，该工具无需设备，技术专业知识不受限制，对发达国家和欠发达国家都有用。这种方法产生了一个非常强大的诊断平台，将补充现有的分析，提供对广泛的疾病的诊断，并能够利用额外的基于酶免疫分析的内容。这项提议是对新兴微阵列技术的有效整合，并将为未来的血清学检测系统奠定基础。

DESCRIPTION (provided by applicant): We aim to develop a fully integrated microfluidic immunodiagnostic platform utilizing a novel air-liquid cavity acoustic transducer (ALCAT) technology pioneered in the lab of Dr Abe Lee. The diagnosis platform is based on the well-established clinical diagnostic tool, the enzyme immunoassay (eg ELISA). ELISAs are an integral platform for clinical diagnosis because they are rapid, robust, and provide results that are highly sensitive and specific. Additionally, ELISAs rely on the inherent humoral immune response of the host to produce antibodies indicative of pathological status. In this proposal, we will form a strategic partnership of expertise in protein microarrays (Antigen Discovery, Inc. and Dr Phil Felgner) and in microfluidics (Dr Abe Lee) to develop a point-of-care diagnostic system consisting of a cost-effective disposable microfluidic plastic chip and a portable analyzer that can be driven by a mobile computing device for rapid serological evaluation of Melioidosis. By synergistically incorporating novel ALCAT-based approaches into these arrays we will have the versatility and power to reduce enzymatic development times, limit production costs, and fully integrate all components into a disposable assay. From previous NIH/NIAID funding we have discovered serodiagnostic antigens for 12 NIAID priority agents (Brucella melitensis, Francisella tularensis, Burkholderia pseudomallei, Coxiella burnetii, Salmonella enterica typhi, Rickettsia rickettsii, Orientia tsutsugamushi, Toxoplasma gondii, M. tuberculosis, variola major related pox viruses, Dengue, and West Nile). We have also identified serodiagnostic antigens for non-biodefense targets including: Bartonella henselae, Borrelia burgdorferi, Hepatitis B Virus, Chlamydia, Plasmodium falciparum, Human papillomavirus, and Herpes simplex virus. This approach will leverage our existing content of serodiagnostic antigens (and appropriate banked sera collection) to produce a multiplex device that will contain multiple antigens from numerous pathogens and enable clinicians to determine which pathogen an individual was exposed to. We believe the platform takes advantage of this wealth of content, and marries it to a versatile ALCAT-based microfluidics approach to produce a true lab-on-a- chip tool that is equipment free, technical expertise unrestricted, and useful for both developed and undeveloped countries. This approach produces a an immensely powerful diagnostic platform that will compliment current assays, provide diagnosis for a broad range of diseases, and is capable of utilizing additional enzyme immunoassay-based content. This proposal is an effective integration of emerging microarray technologies and will set the stage for future serological detection systems.