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: 7277291
• 负责人: Kelly J. Henrickson
• 金额: $ 159.72万
• 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/7277291
• 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

DESCRIPTION (provided by applicant): 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 PCR amplification 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.

描述（由申请人提供）：世界各地的恐怖袭击需要社会对这些不可预测和致命的事件进行强有力的辩护。 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）。严重关注的另一个特工是流感（流感）（“ C”类）。流感A和B病毒每年杀死数十万个全世界，使社会造成数十亿至十亿美元的发病率和社会破坏。对流感的鸟一传播以及人类对人类传播的潜在适应性存在着其他关注。恐怖分子可以利用流感的抗原柔韧性，工程师“改变”了能够引起全球大流行病的更具毒性的菌株。 当前的诊断测定针对流感A（H1N1和H3N2）和FLU B的常见人分离株，但无法检测流感A（16 ha类型，9种N类型）的所有禽类抗原品种。我们的实验室开创了一种同时检测多种病原体的灵活，快速，敏感和特定的方法。我们的多重PCR-EHA测试（Hexaplex，Pneumoplex，Adenoplex，Sars/Coronaplex等）在世界范围内广泛使用。我们最近开发了两种生物包装测定，它们检测到许多（15）个类别“ A”代理。但是，现在的新型DNA检测方法（电子微阵列）和核酸纯化现在可以开发单个“护理点”设备，从而可以增强多路复用测定的速度，灵活性，吞吐量和成本有效性。该应用的具体目的是：1）化学，我们将选择，优化和整合样品制备化学，多重PCR扩增系统和电子微阵列方法，以检测流感A/B的所有抗原型变异（流血流感）（流感流感）类别的大多数“ Bioterrormism Agents Applyerm Agents（Bioterrorism Agent）”（Biotorrism Agent Asseption（Bioterrism Assect）的设备（Bioterrism Assept）（生物抗性）的设备（分开）设备。 2）可行性，使用先前收集的临床样本在小型临床前研究中确定3个模块的性能特征； 3）自动化，微型化，整合和简化SA＃1的化学和机制，以创建3个模块，然后是完全集成的单个设备（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）与非分子方法相比，新开发的测定法的临床灵敏度和特异性以及我们对先前收集的样品的1200个未综合分子测定（PCR-EHA）相比，新开发的测定法的临床灵敏度和特异性。这些新测定在一个集成的单个设备中，可以在门诊环境中1-2小时内进行具有成本效益的护理点诊断。这些目标对最近的NIAID宣布高优先级流感（NOT-AI-05-013）和生物威胁（RFA AI-05-019）研究敏感。