Point of care microfluidic diagnosis system for rapid and sensitive detection of

描述（由申请人提供）：本提案的目的是将基因组学、微流体、DNA和RNA扩增、微阵列、分析仪器开发和聚合物微加工专业知识相结合，以开发一种革命性的医疗诊断器械，用于快速多参数检测呼吸道临床样本中发现的最相关病毒。病毒性呼吸道感染（VRTI）是当今最常见的疾病（Fendrick等人，2003年），仅在美国每年就花费超过500亿美元。据估计，有十几种不同的病毒感染人类上呼吸道，但目前基于病毒抗原的诊断方法往往缺乏所需的灵敏度，而基于培养的方法非常缓慢。大学拉瓦尔及其合作者打算将他们现有的诊断技术整合到一个单一的微流控向心装置（MCD）或芯片实验室中，用于检测VRTI-MCD。VRTI-MCD将自动（i）处理鼻咽拭子样本，（ii）以足够纯的形式提取病毒RNA/DNA，（iii）扩增和标记核酸，（iv）处理标记扩增子，（v）在低密度微阵列上杂交扩增子，（vi）读取和解释数据，以在一小时内提供临床有用的结果，以及（vii）将结果实时传达给健康信息系统。VRTI-MCD将是一种一次性聚合物器械，将封装所有必要的试剂。一次性用品将由便携式和易于使用的光机械仪器处理，实现自动VRTI诊断。17种病毒，包括甲型流感（如禽流感和猪流感）、B流感、SARS冠状病毒、副流感病毒和引起大多数VRTI的腺病毒沿着关键的抗病毒耐药标志物，将使用单一VRTI-MCD检测。这种新型的诊断平台将使用在Centre de Recherch <$en Eschertiologie（CRI）收集的大量临床样本中提供的鼻咽拭子样本进行开发和优化。到NIH支持的第五年结束时，我们将使用人类临床样本验证集成VRTI-MCD，便携式仪器将准备好在全球范围内进行即时诊断的临床测试。此后，在两年内，产品开发和卫生当局（FDA，加拿大卫生部等）的批准。将由一个商业合作伙伴负责。这一新的技术平台将导致病毒性呼吸道感染的即时诊断测试（POCT）的空前扩展，同时在不到一小时的时间内提供快速而经济的分子诊断结果，而不是目前的2-3天。这将改善医疗反应，使人们能够及时使用适当的抗病毒药物，并确保只有在适当的时候才采取快速隔离措施。这些POCT将帮助公共卫生当局为基于实验室和基于现场的应用确定新的操作策略。

DESCRIPTION (provided by applicant): The objective of this proposal is to merge genomics, microfluidics, DNA and RNA amplification, microarrays, analytical instrumentation development and polymer microfabrication expertises to develop a revolutionary medical diagnostic device for the rapid multiparametric detection of the most relevant viruses found in respiratory tract clinical samples. Viral respiratory tract infection (VRTI) is the most common illness today (Fendrick et al., 2003), and costs over $50 billion yearly in the US alone. It is estimated that more than a dozen different viruses infect human upper airways, yet current diagnostic methods based on viral antigens often lack the required sensitivity, while culture-based methods are extremely slow. Universit¿ Laval with its collaborators intends to integrate their already existing diagnostic technologies into a single Microfluidic Centripetal Device (MCD), or Lab-on-a-Chip, for the detection of VRTIs (VRTI-MCD). The VRTI-MCD will automatically (i) process nasopharyngeal swab samples, (ii) extract viral RNA/DNA in a sufficiently pure form, (iii) amplify and label nucleic acids, (iv) process labelled-amplicons, (v) hybridize amplicons on low density microarrays, (vi) read and interpret the data to provide a clinically useful result within one hour, and (vii) communicate results in real-time to health information systems. The VRTI-MCD will be a disposable polymer-based device that will encase all necessary reagents. The disposable will be processed by a portable and easy-to-use opto-mechanical instrument enabling automated VRTI diagnostics. Seventeen viruses, including influenza A (such as avian flu and swine flu), influenza B, SARS coronavirus, parainfluenza, and adenovirus responsible for the majority of VRTI along with key antiviral resistance markers will be detectable using a single VRTI-MCD. This novel diagnostic platform will be developed and optimized using nasopharyngeal swab specimens available in the extensive clinical samples collection at the Centre de Recherch¿ en Infectiologie (CRI). By the end of year-five support by NIH, we will have validated the integrated VRTI-MCD using human clinical samples and the portable instrument will be ready for worldwide clinical testing for diagnostics at the point-of-care. Thereafter, within two years, product development and approval by health authorities (FDA, Health Canada, etc.) will be undertaken by a commercial partner. This new technological platform will result in an unprecedented expansion of point-of-care diagnostic tests (POCTs) for viral respiratory tract infections, while providing rapid yet economical molecular diagnostics results in less than one hour instead of the current 2-3 days. This will improve medical response, allowing for the timely use of proper antiviral drugs and to ensure that rapid quarantine measures only be used when appropriate. These POCTs will help public health authorities to define new operational strategies for both lab-based and field-based applications.