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、乙型流感、副流感(1-3型)、人类呼吸道合胞病毒(RSV)、腺病毒(A-f型)、SARS冠状病毒、A组链球菌、B组链球菌、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)。建议采用基于阵列的呼吸系统疾病诊断方法，因为根据患者的临床症状或放射学很难区分病毒感染和细菌感染，而且众所周知，单一抗体测试容易出现假阳性。提出了一种竞争性免疫分析，以防止夹心免疫分析阵列形式中的检测器抗体交叉反应；夹心免疫分析被保留为实现第一阶段里程碑的应急措施。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，并在获取样本后20分钟内在诊所或现场智能地使用昂贵和/或有限数量的抗生素或抗病毒疗法。我们将利用Akonni从Argonne国家实验室获得的独家许可知识产权组合；从Quidel Corporation获得的专利抗体、样品制备试剂和技术；Argonne国家实验室的蛋白质阵列专业知识；以及Mary医院的小公司的临床实践，来实现这些目标。第一阶段项目的具体目标是：在Akonni的流体发展模型子电路内开发和实施样品制备和多重免疫分析程序；确定目标抗原与竞争蛋白质阵列中Quidel抗体的交叉反应；确定12种病原体在磷酸盐缓冲盐水和改良的鼻拭子、洗液或痰中的检测和交叉反应限度；并确定第二阶段集成射流色谱柱开发、生产和测试的工艺流程和仪器要求。项目里程碑的成功完成将为继续进行第二阶段提案提供充分的理由，其中的具体目标将是：将样品制备和蛋白质阵列功能集成到一个独立的、集成的、注射成型的测试色谱盒中；测试和评估测试色谱盒中试剂的稳定性和寿命；将阵列决策逻辑编码到用户友好的软件中；根据510K提交指南，测试和评估整齐和修改后的鼻拭子、喉咙拭子、鼻腔冲洗和/或痰临床样本的集成原型；并与Quidel合作开始扩大墨盒生产。 呼吸蛋白阵列诊断试剂盒将代表一种低成本、简单的诊断工具，用于识别感染呼吸道病原体的个人。这里开发的底层平台同样将在疾病监测的许多领域得到广泛应用，重点是使用点应用程序。

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.