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

用于检测已知和新出现的呼吸道病毒的 POC 分子方法。

• 批准号: 7743639
• 负责人: JOHN Charles GERDES
• 金额: $ 44.41万
• 依托单位: MICRONICS, INC.
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-01 至 2011-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7743639
• 关键词: Adenoviruses; Antibiotics; Area; Awareness; Bacteria; Bacterial Infections; Biological Assay; Clinic; Clinical; Clinical Research; Clinics and Hospitals; Communities; Consensus; Conserved Sequence; Coronavirus; DNA; Data; Data Set; Detection; Development; Devices; Diagnosis; Diagnostic; Disease; Disease Outbreaks; Early Diagnosis; Economic Burden; Epidemic; Epidemiologic Studies; Etiology; Evaluation; Family; Funding; Genomics; Goals; Health; Health Care Costs; Health Personnel; Heating; Herpesviridae; Hospitalization; Hospitals; Human; Hybrids; Individual; Infection; Infectious Agent; Influenza; Institutes; Lead; Lung diseases; Magnesium; Marketing; Metapneumovirus; Methods; Microfluidic Microchips; Microfluidics; Molecular; Nose; Nucleic Acids; Oligonucleotide Primers; Outcome; Papillomavirus; Parainfluenza; Paramyxovirus; Pathogen detection; Patients; Pediatric Hospitals; Phase; Physicians; Physicians' Offices; Plasmids; Prevention; Prevention strategy; Protocols documentation; Public Health; Reaction; Reading; Reagent; Research; Resources; Respiratory Tract Infections; Respiratory syncytial virus; Retroviridae; SARS coronavirus; Sampling; Screening procedure; Sensitivity and Specificity; Site; Solid; Specificity; Specimen; Swab; Symptoms; System; Techniques; Technology; Testing; Time; United States National Institutes of Health; Universities; Variant; Viral; Virus; Washington; World Health; base; cost; design; disease diagnosis; empowered; experience; improved; innovation; innovative technologies; instrument; member; novel; novel diagnostics; pathogen; point of care; point-of-care diagnostics; prevent; prototype; public health relevance; respiratory; respiratory virus; tool; translational health science; treatment strategy; user-friendly; viral DNA; virology

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.

描述（由申请人提供）：仅在美国，由于呼吸道感染而导致的治疗和住院费用超过400亿美元。呼吸道疾病对人类健康的影响更大，涉及呼吸道病毒的全球流行病的潜力很高。目前，最敏感和最全面的测定无法检测到至少一半具有明显呼吸症状的人的临床样本中的病毒病原体。这些测定中的大多数都是昂贵的，并且不打算用于护理点。在此I阶段应用中，我们建议将两种创新的平台技术结合：我们的分子方法，使用独特的共识分级混合杂种寡核苷酸引物（CodeHops），通过PCR扩增来识别病原体的新成员，并具有完全集成的试剂，包含我们目前在商业开发中进行的快速和准确测试的微流体墨盒。这将提供有史以来的首个基于核酸测定的基于核酸测定，具有成本效益且可靠的原型分子平台，适用于诊断和监视。这种独特的技术组合的预期商业用途是一种筛查方法，可检测未检测到已知病毒的标本中的病毒，以及对新兴感染的全球监测和流行病学研究。我们的目标是通过针对两个构成巨大公共卫生负担的两个突出的呼吸病毒家族，腺病毒和副氟藻病毒来证明这种诊断工具的可行性。具体目的是（1）开发和验证实时CodeHop PCR分析，以基于腺病毒和帕托型病毒家族成员的广泛检测（2）将这些测定器转移到微流体墨盒中，以实现快速，易于廉价且易于 - 易于的 - 在护理点上使用呼吸道病毒的检测； （3）验证来自每个家庭的十个临床分离株的Codehop codehop检测。实现这些目标将支持II期应用，该应用将集中于基于每个病毒家族的CodeHop扩增子的新型病毒菌株特异性固相（VISP）测试，以快速区分PCR反应中被扩增的病毒物种，并且关于将测定的集成到基于微流体的检测系统中。第二阶段的发展还将扩大靶向呼吸道病毒家族的靶向小组，以包括所有引起呼吸系统疾病的病毒。最终产品将包括一次性的一次性微流体墨盒，热稳定剂和便携式仪器（将在II期开发期间提供β系统），以操作完全集成的墨盒并提供读出。即使病原体病原体未知，该基于墨盒的系统也能够在护理点上检测病毒呼吸道病原体。我们预计，这种快速且易于使用的诊断产品的开发将为小型诊所和医院的医生提供急需的工具，以全面测试治疗点的呼吸道疾病。这可能会对我们的监视能力以及预防和治疗策略产生重大影响，并最终改善人类健康。公共卫生相关性：小型医院，诊所和医师办公室中患有呼吸道症状的患者的广泛核酸测定测试是不可能的。我们提出了基于综合且负担得起的微流体系统系统的开发，即使病原体病原体先前未知，该系统也具有独特的能力检测病毒呼吸道病原体。预计该系统将产生更高的诊断率，并允许发现新兴的流行病威胁。该系统将有可能通过立即针对病毒定制治疗，减少抗生素使用和不必要的测试，并减少住院的数量和时间来改善患者预后。它将提供必要的数据来管理爆发并最大程度地减少社区影响。由于呼吸道疾病是如此突出，这显然有可能大大降低医疗费用并大大改善公共卫生。