Rapid point-of-care diagnostic for bioterrorism "A" agents and Pandemic influenza

生物恐怖主义“A”制剂和大流行性流感的快速护理点诊断

• 批准号: 7489862
• 负责人: Kelly J. Henrickson
• 金额: $ 156.11万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-01 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7489862
• 关键词: Adenoviruses; Affect; Ambulatory Care Facilities; Animals; Anthrax disease; Antigenic Variation; Automation; Avian Influenza; Bacillus anthracis; Bacterial Pneumonia; Bathing; Benchmarking; Biological; Biological Assay; Bioterrorism; Birds; Botulinum Toxins; Botulism; Capital; Categories; Cercopithecine Herpesvirus 1; Characteristics; Chemistry; Chickenpox; Clinical; Clinical Sensitivity; Clostridium; Cyclic GMP; DNA; Detection; Development; Devices; Diagnosis; Diagnostic; Disruption; Dose; Drug Formulations; Electronics; Engineering; Ensure; Epidemic; Equipment; Event; Francisella; Francisella tularensis; Funding; Genetic; Genome; Goals; Gold; Grant; Heating; Human; Human Resources; Individual; Infectious Agent; Influenza; Investigation; Investments; Knowledge; Laboratories; Learning; Legal patent; Life; Link; Liquid substance; Mechanics; Methods; Molecular; Molecular Diagnosis; Morbidity - disease rate; National Institute of Allergy and Infectious Disease; Numbers; Outpatients; Parasites; Patients; Performance; Plague; Pliability; Polymerase Chain Reaction; Population; Preparation; Process; RNA Viruses; Reaction; Reagent; Research; Research Personnel; Sampling; Sensitivity and Specificity; Sepsis; Serotyping; Serum; Severe Acute Respiratory Syndrome; Skin; Smallpox; Societies; Specimen; Speed; Sputum; Standards of Weights and Measures; Statistically Significant; Structure; Swab; System; Taxon; Techniques; Testing; Time; Tularemia; United States Food and Drug Administration; United States National Institutes of Health; Variant; Viral; Viral Hemorrhagic Fevers; Virulent; Virus; Water; Work; Yersinia pestis; biothreat; commercialization; cost; cost effective; cost effectiveness; experience; flu; human coronavirus; influenza epidemic; instrument; killings; miniaturize; multiplex detection; nucleic acid purification; pandemic disease; pandemic influenza; pathogen; point of care; preclinical study; programs; prototype; research study; respiratory virus

The terrorist attacks around the world necessitate society's continued investment in a strong defense against these unpredictable and deadly events. Infectious agents identified to pose the greatest potential threat (Category "A" agents) include Variola major (smallpox), Bacillus anthracis (anthrax), Yersinia pestis (plague), Clostridium botulinum toxin (botulism), Francisella tularensis (tularaemia), and a group of RNA viruses that cause hemorrhagic fevers (VHFs). Another agent of grave concern is influenza (Flu) (Category "C"). Flu A and B viruses kill hundreds of thousands each year world wide and cost society tens to hundreds of billion dollars in morbidity and societal disruption. Additional concern exists over bird-to-human spread of Flu and the potential adaptation for human-to-human spread. Terrorist could take advantage of Flu's antigenic flexibility and engineer "shifted" more virulent strains capable of causing worldwide pandemics. Current diagnostic assays are directed to the common human isolates of Flu A (H1N1 and H3N2) and Flu B, but no assay is available to detect all of the avian antigenic varieties of Flu A (16 HA types, 9 N types). Our laboratory has pioneered a flexible, rapid, sensitive, and specific method of simultaneously detecting multiple pathogens. Our multiplex PCR-EHA tests (Hexaplex, Pneumoplex, Adenoplex, SARS/Coronaplex, etc.) are used widely around the world. We have recently developed two BioTplex assays that detect many (15) category "A" agents. However, new methods of amplified DNA detection (electronic microarrays) and nucleic acid purification now allow for the development of a single "point-of-care" device that may enhance the speed, flexibility, throughput, and cost effectiveness of multiplex assays. The Specific Aims of this application are: 1) Chemistry, we will select, optimize, and integrate sample preparation chemistry, multiplex PCRamplification systems, and electronic microarray methods to detect all antigenic variants of Flu A/B (pandemic Flu assay) and the majority of category "A" bioterrorism agents (BioT assay) in an open platform mode (separate equipment for each function); 2) Feasibility, to determine performance characteristics of the 3 modules in small pre-clinical studies using previously collected clinical samples; 3) Automation, miniaturize, integrate, and simplify the chemistry and mechanisms of SA#1 to create 3 modules and then a fully integrated single device (prototype); 4)Process development, ensure manufacturability at a reasonable cost (e.g., critical reagents, specifications, test methods, suppliers, stability, and formulation into components); 5) Pre-Clinical study, test the clinical sensitivity and specificity of the newly developed assays, compared to non-molecular methods and our "gold standard" not integrated molecular assays (PCR-EHA) on 1200 previously collected samples. These new assays, in an integrated single device, may allow cost effective, point-of-care diagnosis within 1-2 hrs in an outpatient setting. These goals are responsive to a recent NIAID announcement for High- Priority influenza (NOT-AI-05-013) and bio-threat (RFA AI-05-019) research.

恐怖袭击世界各地的袭击需要社会继续投资于强有力的辩护 反对这些不可预测和致命的事件。鉴定出具有最大潜力的传染剂 威胁（类别“ A”代理）包括Variola Major（天花），炭疽芽孢杆菌（炭疽芽孢杆菌），鼠疫 （瘟疫），肉毒梭菌毒素（肉毒杆菌），弗朗西斯氏菌（Tularemia）和一组RNA 引起出血发烧的病毒（VHFS）。严重关注的另一个特工是流感（流感）（类别 “ C”）。流感A和B病毒每年杀死数十万个全世界，使社会成千上万 十亿美元发病率和社会破坏。对于鸟瞰者的传播还存在其他问题 流感和人对人类传播的潜在适应。恐怖分子可以利用流感 抗原灵活性和工程师“转移”了能够引起全球大流行的更具毒性的菌株。 当前的诊断测定针对流感A（H1N1和H3N2）和流感B的常见人分离株， 但是，无法检测流感A的所有鸟类抗原品种（16公顷，9种N类型）。我们的 实验室已经开创了一种同时检测多个的灵活，快速，敏感和特定的方法 病原体。我们的多重PCR-EHA测试（Hexaplex，Pneumoplex，Adenoplex，Sars/Coronaplex等）是 在世界各地广泛使用。我们最近开发了两个检测到许多的生物质体测定（15） 类别“ A”代理。但是，新的放大DNA检测方法（电子微阵列）和核酸 酸性纯化现在允许开发单个“护理点”设备，该设备可能会提高速度， 多重测定的灵活性，吞吐量和成本效益。该应用程序的具体目的是：1） 化学，我们将选择，优化和整合样品制备化学，多重pcramplification 系统和电子微阵列方法，用于检测流感A/B的所有抗原变异（大流动测定） 以及大多数类别的“ A”生物恐怖剂（Biot Assay）以开放平台模式（独立的平台模式） 每个功能的设备）； 2）可行性，确定3个模块的性能特征 临床前研究使用先前收集的临床样本； 3）自动化，小型化，整合， 并简化SA＃1的化学和机制创建3个模块，然后是一个完全集成的单个模块 设备（原型）; 4）流程开发，以合理的成本确保生产性（例如，关键 试剂，规格，测试方法，供应商，稳定性以及组件的配方）； 5）临床前 与非分子相比，研究，测试新开发测定的临床灵敏度和特异性 方法和我们的“黄金标准”未整合分子测定（PCR-EHA）先前收集的1200 样品。这些新测定在集成的单个设备中可以允许具有成本效益的护理点诊断 在门诊设置中1-2小时内。这些目标是对最近的NIAID公告的反应 优先型流感（非AI-05-013）和生物威胁（RFA AI-05-019）研究。