Point of Care Instrument for Viral Respiratory Diagnosis

用于病毒呼吸道诊断的护理点仪器

• 批准号: 6666700
• 负责人: Jeffrey T. Ives
• 金额: $ 48.71万
• 依托单位: XTRANA, INC.
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-30 至 2004-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6666700
• 关键词: Orthomyxoviridae; clinical research; consumable /disposable biomedical equipment; cooperative study; diagnosis design /evaluation; human tissue; parainfluenza virus type 1; parainfluenza virus type 2; polymerase chain reaction; respiratory disorder diagnosis; respiratory infections; respiratory syncytial virus; respiratory virus; rhinovirus

DESCRIPTION (provided by applicant): The common respiratory viruses (influenza, parainfluenza, respiratory syncytial virus or RSV, and rhinovirus) cause millions of cases of upper and lower respiratory tract disease every year. The spectrum of illness induced directly by the viruses is very broad, ranging from mild colds and simple ear infections to more serious and sometimes life-threatening diseases such as bronchiolitis, pneumonia, and asthma. In spite of the importance of diagnosing and treating these viral infections, current diagnostic methods of physical exam or rapid immunoassays or culture in reference laboratories do not provide the accuracy, sensitivity, or timely information that the primary care physician needs for treatment. Antiviral therapeutics are becoming increasingly viable, but they require accurate identification of the infectious virus(es). While sensitive, newer assays and instrumentation based on nucleic acid technologies are not cost effective or practical for point of care in a physician's office. To provide a cost effective, point of care system for diagnosing respiratory viruses, this application describes an integrated and unique device that will begin with a nasal wash sample and use nucleic acid-based technologies to accurately identify eight respiratory viruses: RSV A and B; parainfluenza 1, 2, and 3; influenza A and B; and rhinovirus. After loading the sample, proprietary steps of sample preparation, RNA extraction, multiplexed amplification, and detection by lateral flow strips will be automatically performed. These processing steps will be directed on a disposable, microfluidic card that both minimizes the reagent costs and provides an enclosed container for convenient, biosafe disposal. To develop this diagnostic device, a series of technical milestones will be accomplished. First, a novel method of multiplexed target amplification will be optimized on respiratory viral targets. Second, nasal wash samples will be directly prepared for multiplexed amplification by flow through extraction and unique solid phase analysis. Sample preparation and amplification will be managed by developing disposable, laminated microfluidics cards with channels, reagents, and valves. Using a protoype workstation, the described system will be tested with patient samples and spiked samples to verify the utility, specificity, sensitivity, and reproducibility of the system.

描述（申请人提供）：常见呼吸道病毒（流感、副流感、呼吸道合胞病毒或RSV、鼻病毒）每年引起数百万例上下呼吸道疾病。 由病毒直接引起的疾病范围非常广泛，从轻微的感冒和简单的耳部感染到更严重的、有时危及生命的疾病，如细支气管炎、肺炎和哮喘。 尽管诊断和治疗这些病毒感染很重要，但目前的体检诊断方法或参考实验室的快速免疫测定或培养方法无法提供初级保健医生治疗所需的准确性、敏感性或及时信息。 抗病毒治疗变得越来越可行，但它们需要准确识别传染性病毒。 虽然基于核酸技术的新型检测和仪器很灵敏，但对于医生办公室的护理点而言并不具有成本效益或不实用。 为了提供一种具有成本效益的护理点系统来诊断呼吸道病毒，该应用描述了一种集成且独特的设备，该设备将从鼻洗样本开始，并使用基于核酸的技术来准确识别八种呼吸道病毒：RSV A 和 B；副流感 1、2 和 3 型；甲型和乙型流感；和鼻病毒。 加载样品后，将自动执行样品制备、RNA 提取、多重扩增和侧流条检测的专有步骤。 这些处理步骤将在一次性微流体卡上进行，该卡既可以最大限度地降低试剂成本，又可以提供封闭的容器，以方便、生物安全地处理。 为了开发这种诊断设备，将实现一系列技术里程碑。 首先，将针对呼吸道病毒靶标优化多重靶标扩增的新方法。 其次，鼻洗液样品将通过流通萃取和独特的固相分析直接制备用于多重扩增。 将通过开发带有通道、试剂和阀门的一次性层压微流体卡来管理样品制备和扩增。 使用原型工作站，将使用患者样本和加标样本对所描述的系统进行测试，以验证系统的实用性、特异性、灵敏度和再现性。