Integrated, on-chip RT-MME-PCR for influenza diagnostics; Phase 1 = sample prepar

描述（由申请方提供）：流感病毒是属于正粘病毒科的高度传染性、节段性负义RNA病毒，每年造成约114，000例住院和20，000例死亡。美国食品和药物管理局（FDA）批准了几种药物用于治疗流感感染，但成功的治疗取决于在疾病的前48小时内服用这些药物。最近在东亚爆发的H5 N1禽流感进一步加剧了人们对另一次大流行的担忧，并强调了在监测和诊断应用中需要全面的毒株亚型分型。因此，本申请的目的是基于Akonni的凝胶元件阵列技术开发用于临床标本中的流感诊断的集成的样品-应答PCR阵列。第1阶段的具体目标是开发和实施用于检测盒上流感RNA样本制备和检测的内标;开发用于病毒RNA分离和纯化的样本制备子回路;以及评估流感样本制备效率和改良临床样本的有效性。在第一阶段的成功将导致第二阶段的建议，具体目标将是建立流感逆转录酶，多重，微阵列增强聚合酶链反应将样品制备和RT-MME-PCR阵列功能集成到塑料测试盒中;并在纯净和改良鼻拭子、咽喉拭子、鼻灌洗液和/或痰临床样本中的广泛流感样本组上测试和评估集成原型，为Quidel Corporation的510 K提交做准备。第2阶段结束时的性能目标是在103 TCID 50 mL-1鼻拭子、咽拭子或鼻灌洗液的输入目标浓度下，在2小时或更短时间内获得样本至应答结果。在修正后的临床样本中成功展示集成的多重RT-MME-PCR芯片将为实施Akonni的便携式监测和诊断系统奠定基础，该系统用于RNA病毒，上呼吸道病原体和感染的快速，痕量检测和表征。 流感病毒具有高度传染性，仅在美国每年就造成约114，000人住院治疗和20，000人死亡。东亚不断出现新的病毒株（如甲型流感H5 N1），突出表明需要迅速、全面的监测和诊断工具。Akonni的技术和产品解决方案提供了一个简单的路径，在一个集成的，样品到答案的测试盒和现场便携式仪器的微量RNA病毒和传染病的诊断检测。

DESCRIPTION (provided by applicant): Influenza viruses are highly contagious, segmented negative-sense RNA viruses that belong to the family Orthomyxoviridae, causing approximately 114,000 hospitalizations and 20,000 deaths annually. Several drugs are approved by the US Food and Drug Administration (FDA) for treating influenza infections, but successful treatment depends on administering these drugs within the first 48 h of illness. The recent outbreak of avian influenza H5N1 in East Asia has further fueled concern over another pandemic and emphasizes the need for comprehensive strain sub-typing in surveillance and diagnostic applications. The objective of this Application is therefore to develop an integrated, sample-to-answer PCR array for influenza diagnostics in clinical specimens based on Akonni's gel element array technology. Specific aims in Phase 1 are to develop and implement an internal standard for on-cartridge influenza RNA sample preparation and detection; develop a sample preparation sub-circuit for viral RNA isolation and purification; and assess influenza sample preparation efficiency and efficacy in amended clinical samples. Success in Phase 1 will result in a Phase 2 proposal where the specific aims will be to establish influenza reverse transcriptase, multiplexed, microarray-enhanced polymerase chain reaction (RT-MME-PCR) amplification protocols on the Akonni gel element array platform; integrate the sample preparation and RT-MME-PCR array functions into a plastic test cartridge; and test and evaluate the integrated prototype on an extensive panel of influenza in neat and amended nasal swabs, throat swabs, nasal lavage and/or sputum clinical samples in preparation for 510K submission with Quidel Corporation. The performance goal at the end of Phase 2 is a sample-to-answer result in 2 hours or less at an input target concentration of 103 TCID50 mL-1 nasal swab, throat swab, or nasal lavage. Successful demonstration of an integrated, multiplexed RT-MME-PCR chip in amended clinical samples will lay the foundation for implementing Akonni's portable surveillance and diagnostic system for the rapid, trace detection and characterization of RNA viruses, upper respiratory pathogens and infections. Influenza viruses are highly contagious and cause approximately 114,000 hospitalizations and 20,000 deaths annually in the U.S. alone. The continued emergence of new strains (such as influenza A H5N1) in East Asia highlights the need for rapid, comprehensive surveillance and diagnostic tools. Akonni's technology and product solutions provide a straightforward path for diagnostic detection of trace RNA viruses and infectious diseases in an integrated, sample-to-answer test cartridge and field portable instrument.