POC molecular method for detection of known and emerging respiratory viruses.

描述（由申请人提供）：仅在美国，由于呼吸道感染导致的治疗和住院治疗每年花费超过400亿美元。呼吸道疾病对人类健康的影响在世界范围内甚至更大，涉及呼吸道病毒的全球流行病的可能性很高。目前，最敏感和最全面的检测方法无法在至少一半来自具有明确呼吸道症状的个体的临床样本中检测到病毒病原体。这些测定中的大多数是昂贵的，并且不打算用于护理点使用。在第一阶段的申请中，我们建议将联合收割机两种创新平台技术结合起来：我们的分子方法，使用独特的共识简并杂交寡核苷酸引物池（CODEHOPs）通过PCR扩增识别病原体家族的新成员，以及完全集成的，包含微流体试剂盒的试剂，能够进行快速准确的检测，我们目前正在商业开发中。这将提供第一个基于核酸测定的、具有成本效益的和稳健的原型分子平台，适用于床旁诊断和监测。这种独特的技术组合的预期商业用途是作为一种筛查方法，用于检测尚未检测到已知病毒的标本中的病毒，以及用于对新出现的感染进行全球监测和流行病学研究。我们的目标是通过靶向两个主要的呼吸道病毒家族（腺病毒和副流感病毒）来证明这种诊断工具的可行性，这两个家族构成了重大的公共卫生负担。具体目的是（1）开发和验证用于广泛检测腺病毒和副粘病毒家族成员的实时CODEHOP PCR检测试剂盒（2）将这些检测试剂盒转移到微流体检测盒中，以便在床旁快速、廉价和易于使用地检测呼吸道病毒;（3）验证CODEHOP检测试剂盒对每个家族10种临床分离株的检测。实现这些目标将支持II期申请，该申请将侧重于开发基于每个病毒家族的CODEHOP扩增子的新型病毒株特异性固相（VISP）检测，以快速区分PCR反应中扩增的病毒种类，并将该检测方法整合到基于微流体的检测系统中。第二阶段的发展还将扩大呼吸道病毒家族的目标小组，以包括所有引起呼吸道疾病的病毒。最终产品将包括一次性使用的微流体检测盒、热稳定剂和便携式仪器（在第二阶段开发期间将提供beta系统），以操作完全集成的检测盒并提供读数。这种基于细菌的系统将能够在护理点检测病毒性呼吸道病原体，即使致病病原体是未知的。我们希望这种快速易用的诊断产品的开发将为小型诊所和医院的医生提供一种急需的工具，用于在护理点对呼吸系统疾病进行全面检测。这可能对我们的监测能力以及预防和治疗战略产生重大影响，并最终改善人类健康。公共卫生相关性：在小医院、诊所和医生办公室对有呼吸道症状的患者进行广泛的核酸检测，今天是不可能的。我们建议开发一种全面的、负担得起的基于微流体的即时检测系统，该系统能够独特地检测病毒性呼吸道病原体，即使致病病原体以前是未知的。预计这一系统将提高诊断率，并能发现新出现的流行病威胁。该系统将有可能通过立即调整病毒治疗，减少抗生素使用和不必要的测试，以及减少住院次数和次数来改善患者的治疗效果。它将提供必要的数据，以管理疫情并尽量减少社区影响。由于呼吸道疾病如此突出，所有这一切显然都有可能极大地减少保健费用，并大大改善公共卫生。

DESCRIPTION (provided by applicant): Treatments and hospitalizations due to respiratory infections incur costs exceeding $40 billion dollars yearly in the US alone. The impact of respiratory disease on human health is even greater worldwide and the potential of a global epidemic involving respiratory viruses is high. Currently, the most sensitive and comprehensive assays fail to detect a viral pathogen in at least half of clinical samples from individuals with clear respiratory symptoms. Most of these assays are expensive and not intended for point-of-care use. In this Phase I application, we propose to combine two innovative platform technologies: our molecular approach, using a unique pool of consensus-degenerate hybrid oligonucleotide primers (CODEHOPs) to identify novel members of pathogen families by PCR amplification, with a fully integrated, reagent containing microfluidics cartridge capable of rapid and accurate testing that we currently have in commercial development. This will provide a first ever nucleic acid assay based, cost-effective and robust prototype molecular platform suitable for point-of- care diagnosis and surveillance. The intended commercial use of this unique combination of technologies is as a screening method to detect viruses in specimens for which known viruses have not been detected and for global surveillance and epidemiological studies of emerging infections. Our objective is to demonstrate the feasibility of this diagnostic tool by targeting two prominent respiratory virus families, adenoviruses and parainfluenzaviruses, that constitute a significant public health burden. The specific aims are (1) to develop and validate real-time CODEHOP PCR assays for broad-based detection of members of the adenovirus and paramyxovirus families (2) to transfer these assays to microfluidics cartridges enabling fast, inexpensive, and easy-to-use detection of respiratory viruses at point-of-care; and (3) to verify CODEHOP on-cartridge detection for ten clinical isolates from each family. Achieving these aims would support a Phase II application that would focus on the development of a novel viral strain-specific solid-phase (VISP) test based on the CODEHOP amplicons for each virus family to rapidly distinguish viral species amplified in the PCR reaction, and on the integration of the assay into a microfluidics-based detection system. Phase II development would also broaden the targeted panel of respiratory virus families to include all viruses causing respiratory disease. The final product will comprise disposable, single-use microfluidics cartridges, heat-stable agents and a portable instrument (beta system will be available during Phase II development) to operate the fully integrated cartridge and provide the read-out. This cartridge-based system will be capable of detecting viral respiratory pathogens at point-of-care even if the causative pathogen is unknown. We expect that the development of such a rapid and easy-to-use diagnostic product would provide physicians in small clinics and hospitals with a much needed tool for comprehensive testing of respiratory diseases at point-of-care. This could have a large impact on our surveillance capacities as well as prevention and treatment strategies and ultimately improve human health. PUBLIC HEALTH RELEVANCE: Widespread nucleic acid assay testing of patients with respiratory symptoms in small hospitals, clinics and physician's offices is not possible today. We propose the development of a comprehensive and affordable point-of-care microfluidics-based system that is uniquely able to detect viral respiratory pathogens even if the causative pathogen was previously unknown. This system is expected to yield higher rates of diagnosis and allow detection of emerging epidemic threats. The system will have the potential to improve patient outcomes by immediate tailoring of the therapy to the virus, reducing antibiotic use and unnecessary testing, and decreasing the number and times of hospitalization. It will provide data necessary to manage outbreaks and minimize community impact. Since respiratory illnesses are so prominent all this clearly has the potential to reduce health-care costs enormously and improve public health significantly.