Multiplex Serodiagnostic Protein Microarray

多重血清诊断蛋白质微阵列

• 批准号: 7458234
• 负责人: PHILIP Louis FELGNER
• 金额: $ 74.98万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-08-01 至 2013-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7458234
• 关键词: Acids; Advanced Development; Agreement; Animal Model; Animals; Antibody Formation; Antigen Targeting; Antigens; Appendix; Arboviruses; Asia; Attenuated; Bacteria; Bioinformatics; Biological Assay; Biology; Bioterrorism; Brucella; Brucella melitensis; Budgets; Burkholderia; Burkholderia mallei; Burkholderia pseudomallei; Businesses; California; Categories; Characteristics; Cloning; Collection; Commit; Communicable Diseases; Contracts; Core Protein; Coxiella; Coxiella burnetii; Custom; Data; Data Set; Databases; Dengue Virus; Developed Countries; Developing Countries; Development; Devices; Diagnosis; Diagnostic; Diagnostic Procedure; Diagnostic tests; Discipline; Disease; Doctor of Medicine; Doctor of Philosophy; Emerging Communicable Diseases; Employee; Epidemic; Epitopes; Equipment; Equipment and supply inventories; Evaluation; Family; Fee-for-Service Plans; Funding; Funding Agency; Genomics; Government Agencies; Grant; Guanosine Monophosphate; Hantavirus; Hawaii; Homologous Protein; Hospitals; Human; Human Resources; Immune response; Immunity; Immunodominant Antigens; Immunology; In Vitro; Individual; Industry; Infection; Infectious Agent; Institutes; Institution; Internet; Laboratories; Lateral; Licensing; Longevity; Maryland; Medical; Methodology; Methods; Military Personnel; Monitor; Nucleic Acids; Numbers; Open Reading Frames; Organism; Parasites; Participant; Pathway interactions; Patients; Performance; Peru; Pilot Projects; Plasmids; Polymerase Chain Reaction; Population; Prevalence; Principal Investigator; Printing; Protein Microchips; Proteins; Proteome; Proteomics; Public Health; Publications; Publishing; Purpose; Reagent; Recombinant Proteins; Recombinants; Research; Research Personnel; Resource Sharing; Resources; Rickettsia; Rickettsia prowazekii; Safety; Salmonella; Salmonella infections; Salmonella typhi; Sampling; Scientist; Screening procedure; Sensitivity and Specificity; Serologic tests; Serotyping; Serum; Services; Site; South American; Study of serum; Subunit Vaccines; Surrogate Endpoint; System; Technology; Testing; Texas; Therapeutic; Universities; Vaccinated; Vaccine Antigen; Vaccines; Validation; Vascular blood supply; Virulence; Virus; Walkers; Western Blotting; Work; base; biodefense; cohort; college; commercialization; cost; design; falls; high throughput analysis; improved; interest; man; microorganism; microorganism antigen; multidisciplinary; pathogen; pre-clinical; preclinical study; professor; programs; prototype; rapid detection; rapid technique; response; volunteer

DESCRIPTION (provided by applicant): We have developed technology that enables rapid, comprehensive and high-throughput analysis of humoral immune responses to infectious disease antigens that is being used for subunit vaccine antigen discovery and to develop serodiagnostic tests. The method is based on printing microarray chips with proteins encoded by infectious microorganisms which cause medically important diseases in humans. Currently, we have expressed and printed more than 14,000 individual proteins from 6 bacteria, 17 viruses, and 2 parasites. The chips are probed with serum from infected patients and healthy controls to identify the dominant antibody responses, to determine the immunodominant antigen profile and identify a set of 10-20 serodiagnostic antigens for each infectious agent. By using a multiplicity of antigens for each infectious agent, serodiagnostic tests with statistically improved sensitivity and specificity can be configured. Here we will fabricate whole proteome microarrays for Coxiella burnetii, Rickettsia prowazekii, Brucella melitensis, Salmonella typhi, and non-homologous proteins from related strains of each agent. The arrays will be probed with serum from well-characterized infected subjects and healthy controls to identify the immunodominant and serodiagnostic antigen sets. We will also fabricate a chip containing 190 proteins from 16 arboviruses and 8 hantaviruses. The ability of this virus chip to distinguish well-characterized sera collected from humans and animals infected with the different viruses will be determined and the results compared with established assays done on the same samples. Deliverables generated from this project include plasmids, customized protein microarrays in a modular multiplex format, purified recombinant proteins and lateral flow (dipstick)/immunostrips devices, and will be accessible from the PSWRCE Protein Microarray Core on a recharge basis. These will be available to RCE investigators, government agencies and to private businesses through licensing agreements. The Protein Microarray Core will also provide chip probing and analysis services. Serodiagnostic antigen sets will be available for licensing to businesses interested in developing serodiagnostic tests. The immunodominant antigen sets, which may contain subunit vaccine antigens, will be available to commercial vaccine developers through licensing agreements. Multiplex serodiagnostic chips will be useful for determining whether military personnel or civilians, who enter a region where these agents are endemic, are exposed to any of them. They will also benefit public health monitoring activities to track annual changes in the prevalence of infections in endemic regions, and for monitoring the blood supply in developing countries. The chips will also be useful for assessing the spread of exposure in a population following a bioterrorism attack.

描述（由申请人提供）：我们开发了一种技术，能够快速、全面和高通量地分析对传染病抗原的体液免疫应答，该技术正用于亚单位疫苗抗原发现和开发血清学诊断试验。该方法基于用感染性微生物编码的蛋白质打印微阵列芯片，所述感染性微生物在人类中引起医学上重要的疾病。目前，我们已经表达和打印了来自6种细菌，17种病毒和2种寄生虫的14，000多个蛋白质。用来自感染患者和健康对照的血清探测芯片，以鉴定显性抗体应答，确定免疫显性抗原谱，并鉴定每种感染因子的一组10-20种血清诊断抗原。通过对每种传染原使用多种抗原，可以配置具有统计学上改进的灵敏度和特异性的血清诊断测试。在这里，我们将制作完整的蛋白质组微阵列的贝氏柯克斯体，普氏立克次体，羊种布氏杆菌，伤寒沙门氏菌，和非同源蛋白质从相关菌株的每一个代理。将用来自充分表征的感染受试者和健康对照的血清探测阵列，以鉴定免疫显性和血清诊断抗原集。我们还将制作一个包含16种虫媒病毒和8种汉坦病毒的190种蛋白质的芯片。将确定该病毒芯片区分从感染不同病毒的人类和动物收集的充分表征的血清的能力，并将结果与对相同样品进行的已建立的测定进行比较。该项目产生的可重复使用的产品包括质粒、模块化多重格式的定制蛋白质微阵列、纯化的重组蛋白和侧流（试纸）/免疫条装置，并将在充电基础上从PSWRCE蛋白质微阵列核心获得。这些将通过许可协议提供给RCE调查人员、政府机构和私营企业。蛋白质微阵列核心还将提供芯片探测和分析服务。对开发血清诊断检测感兴趣的企业将获得血清诊断抗原集的许可。免疫显性抗原组可能含有亚单位疫苗抗原，将通过许可协议提供给商业疫苗开发商。多重血清诊断芯片将有助于确定军事人员或平民，谁进入一个地区，这些代理人流行，暴露于其中任何一个。它们还将有利于公共卫生监测活动，以跟踪流行地区感染流行率的年度变化，并监测发展中国家的血液供应。这些芯片还将用于评估生物恐怖袭击后人群中暴露的传播。