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体免疫诊断平台，利用一种新的气液腔声换能器（ALCAT）技术，在实验室中率先安倍李博士。诊断平台基于成熟的临床诊断工具，酶免疫测定法（例如ELISA）。ELISA是临床诊断的一个不可或缺的平台，因为它们快速、稳健，并提供高度灵敏和特异的结果。此外，ELISA依赖于宿主的固有体液免疫应答来产生指示病理状态的抗体。在这项提案中，我们将形成一个战略伙伴关系的专业知识，在蛋白质微阵列（抗原发现，公司。和Phil Felgner博士）和微流体学（Abe Lee博士）开发了一种即时诊断系统，该系统由具有成本效益的一次性微流体塑料芯片和便携式分析仪组成，该分析仪可以由移动的计算设备驱动，用于类鼻疽的快速血清学评估。通过将基于ALCAT的新方法协同并入这些阵列中，我们将具有多功能性和能力来减少酶的开发时间，限制生产成本，并将所有组件完全集成到一次性检测中。从以前的NIH/NIAID基金中，我们已经发现了12种NIAID优先因子的血清诊断抗原（羊种布氏杆菌、土拉弗朗西斯菌、类鼻疽伯克霍尔德菌、贝氏柯克斯体、伤寒沙门氏菌、立克次体、恙虫病东方体、刚地弓形虫、M.结核病、与痘病毒相关的天花、登革热和西尼罗河病毒）。我们还确定了非生物防御目标的血清诊断抗原，包括：汉赛巴尔通体、伯氏疏螺旋体、B型肝炎病毒、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（和适当的库存血清收集）的内容，以产生一个多重装置，将包含来自许多病原体的多种抗原，并使临床医生能够确定一个人暴露于哪种病原体。我们相信，该平台利用了这一丰富的内容，并将其与基于ALCAT的多功能微流体方法结合起来，生产出一种真正的芯片实验室工具，该工具无需设备，技术专长不受限制，对发达国家和不发达国家都很有用。这种方法产生了一个非常强大的诊断平台，它将补充当前的检测方法，为广泛的疾病提供诊断，并能够利用其他基于酶免疫测定的内容。该建议是新兴的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。 公共卫生相关性：在资源充足的参考实验室进行集中检测，无法充分满足大规模灾害、自然疾病爆发和生物恐怖主义威胁期间对快速诊断的不可预测的需求。在这种紧急情况下，急救人员必须依靠准确的诊断，这些诊断可以快速有效地部署到可能不容易识别感染体征或症状的患者身上。护理点（POC）多重诊断非常适合担任这一重要角色，因为它们高度便携，不需要广泛的培训，并提供快速的样本到回答时间。

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. PUBLIC HEALTH RELEVANCE: Centralized testing in well-resourced reference laboratories cannot adequately address the unpredictable demand for rapid diagnostics during mass disasters, natural disease outbreaks, and bioterrorist threats. During such emergencies, first responders must rely on accurate diagnostics that are quickly and efficiently deployable to patients who may not have easily recognizable signs or symptoms of an infection. Point-of-care (POC) multiplex diagnostics are well-suited to fill this important role as they are highly portable do not require extensive training, and provide prompt sample-to-answer times.