Diagnostic protein array for respiratory infections

描述（申请人提供）：该项目的目标是将Argonne开发的知识产权转化为商业市场，并开发用于临床监测和诊断常见呼吸道感染的蛋白质阵列，包括甲型流感、B型流感、副流感（1-3型）、人呼吸道合胞病毒（RSV）、腺病毒（A-f型）、SARS冠状病毒、A群链球菌、B群链球菌、结核分枝杆菌、肺炎支原体、流感嗜血杆菌、和肺炎葡萄球菌（12种病原体）。推荐使用基于阵列的方法来诊断呼吸道疾病，因为很难根据患者的临床体征或放射学来区分病毒和细菌感染，并且因为单抗体测试非常容易出现假阳性。提出了竞争性免疫测定，以抑制夹心免疫测定阵列形式中的检测器抗体交叉反应性;夹心免疫测定保留作为实现I期里程碑的应急措施。来自蛋白质阵列的决策逻辑和结果将使公共卫生官员能够快速识别感染者，并在样本采集后20分钟内在诊所或现场智能地施用昂贵和/或有限数量的抗生素或抗病毒疗法。我们将利用Akonni从阿贡国家实验室获得的独家许可知识产权组合; Quidel Corporation的专有抗体、样品制备试剂和专有技术;阿贡国家实验室的蛋白质阵列专业知识;以及Little Company of玛丽医院的临床实践来实现这些目标。第1阶段项目的具体目标是：在Akonni的流体开发模型子回路中开发和实施样品制备和多重免疫测定程序;确定竞争性蛋白阵列中靶抗原与Quidel抗体的交叉反应性;确定磷酸盐缓冲盐水和改良鼻拭子、灌洗液或痰液中12种单独病原体的检测限和交叉反应性;并定义第2阶段中集成射流卡盘开发、生产和测试的工艺流程和仪器要求。成功完成项目Milestone将为继续进行第2阶段提案提供充分的理由，其中具体目标将是：将样本制备和蛋白质阵列功能集成到独立的、集成的、注塑成型的检测盒中;检测和评价检测盒内的试剂稳定性和寿命;将阵列决策逻辑编码到用户友好型软件中;根据510 K提交指南，在纯鼻拭子、咽拭子、鼻灌洗液和/或痰液临床样本上测试和评价集成原型;并与Quidel合作，开始测试卡片生产规模扩大。 诊断呼吸道蛋白阵列检测盒将代表一种低成本、简单的诊断工具，用于识别呼吸道病原体感染的个体。本文开发的基础平台同样将在疾病监测的许多领域中找到广泛的应用，重点是使用点应用。

DESCRIPTION (provided by applicant): The objective of this project is to transition Argonne-developed intellectual property into the commercial marketplace, and develop a protein array for clinical surveillance and diagnosis of common respiratory infections, including influenza A, influenza B, parainfluenza (types 1-3), human respiratory syncitial virus (RSV), adenovirus (types A-f), SARS coronavirus, group A Streptococcus, group B Streptococcus, Mycobacterium tuberculosis, Mycoplasma pneumonia, Haemophilus influenza, and Staphylococcus pneumoniae (12 pathogens). An array-based approach to respiratory disease diagnosis is recommended because it is difficult to differentiate between viral and bacterial infections based on clinical signs or radiology of the presenting patient, and because single-antibody tests are notoriously prone to false positives. A competitive immunoassay is proposed to militate against detector antibody cross-reactivity in a sandwich immunoassay array format; the sandwich immunoassay is reserved as a contingency to achieve the Phase 1 milestones. The decision logic and results from the protein array will enable public health officials to rapidly identify infected persons and intelligently administer costly and/or limited quantities of antibiotics or anti-viral therapies either in the clinic or in the field within 20 min of sample acquisition. We will meet these objectives by taking advantage of Akonni's exclusively-licensed intellectual property portfolio acquired from Argonne National Laboratory; proprietary antibodies, sample preparation reagents and know-how from Quidel Corporation; protein array expertise at Argonne National Laboratory; and clinical practice at Little Company of Mary Hospital. Specific Aims for the Phase 1 project are to: develop and implement a sample preparation and multiplexed immunoassay procedure within Akonni's fluidic development model sub-circuits; determine cross- reactivity of target antigens with Quidel antibodies in the competitive protein array; determine limits of detection and cross-reactivity for 12 individual pathogens in phosphate buffered saline and amended nasal swabs, lavage or sputum; and define process flow and instrument requirements for integrated fluidic cartridge development, production and testing in Phase 2. Successful completion of the project Milestones will provide sufficient justification to proceed with a Phase 2 proposal, where the Specific Aims will be to: integrate the sample preparation and protein array functions into a self-contained, integrated, injection-molded test cartridge; test and evaluate reagent stability and longevity within the test cartridge; encode the array decision logic into user-friendly software; test and evaluate the integrated prototype on neat and amended nasal swabs, throat swabs, nasal lavage and/or sputum clinical samples in accordance with 510K submission guidelines; and begin cartridge production scale-up in collaboration with Quidel. The Diagnostic Respiratory Protein Array test cartridge will represent a low-cost, simple diagnostic tool for identifying individuals infected with respiratory pathogens. The underlying platform developed herein will likewise find broad application in many areas of disease surveillance, with an emphasis on point-of-use applications.