Multiplex Serodiagnostic Protein Microarray

多重血清诊断蛋白质微阵列

• 批准号: 8137655
• 负责人: PHILIP Louis FELGNER
• 金额: $ 74.01万
• 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/8137655
• 关键词: Agreement; Animals; Antibody Formation; Antigens; Arboviruses; Bacteria; Biological Assay; Bioterrorism; Brucella melitensis; Businesses; Categories; Cloning; Communicable Diseases; Contracts; Coxiella burnetii; Data Set; Dengue Virus; Developing Countries; Devices; Diagnosis; Diagnostic Procedure; Diagnostic tests; Disease; Emerging Communicable Diseases; Epidemic; Equipment and supply inventories; Evaluation; Family; Government Agencies; Hantavirus; Hospitals; Human; Human Resources; Immune response; Immunodominant Antigens; Individual; Infection; Infectious Agent; Laboratories; Lateral; Licensing; Methodology; Methods; Military Personnel; Monitor; Parasites; Patients; Performance; Plasmids; Population; Prevalence; Printing; Protein Microchips; Proteins; Proteome; Public Health; Recombinant Proteins; Recombinants; Research Personnel; Resources; Rickettsia prowazekii; Salmonella typhi; Sampling; Screening procedure; Sensitivity and Specificity; Serologic tests; Serotyping; Serum; Services; Subunit Vaccines; System; Technology; Testing; Vaccine Antigen; Vaccines; Validation; Vascular blood supply; Virus; base; cohort; design; high throughput analysis; improved; interest; microorganism; microorganism antigen; pathogen; prototype; response

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）：我们开发了技术，可以快速，全面和高通量分析对传染病抗原的体液免疫反应，该反应用于亚基疫苗抗原发现并开发血清诊断测试。该方法基于印刷微阵列芯片，该芯片是由传染性微生物编码的蛋白质，这些蛋白质引起了人类的医学重要疾病。目前，我们从6种细菌，17种病毒和2个寄生虫中表达并印刷了14,000多个单独的蛋白质。用受感染患者和健康对照的血清探测芯片，以鉴定显性抗体反应，以确定免疫主导抗原谱，并确定每种感染剂的10-20种血清诊断抗原。通过为每个感染剂使用多种抗原，可以配置具有统计学上提高灵敏度和特异性的血清诊断测试。在这里，我们将为Coxiella burnetii，立二氏菌，脑状菌，斑疹伤寒沙门氏菌和非同源蛋白质制作全蛋白质组微阵列。阵列将用特征良好的感染受试者和健康对照的血清探测，以鉴定免疫诊断和血清诊断抗原套件。我们还将制造一个芯片，其中包含来自16个arboviruse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人类和感染不同病毒的动物的良好表征的血清的能力，并且与在相同样品上进行的既定测定法相比结果。该项目产生的可交付成果包括质粒，具有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向流动（二金）/免疫物质设备，并且可以从PSWRCE蛋白质微阵列访问。这些将通过许可协议提供给RCE调查人员，政府机构和私人企业。蛋白质微阵列核心还将提供芯片探测和分析服务。血清诊断抗原集将用于有兴趣开发血清诊断测试的企业的许可。可能包含亚基疫苗抗原的免疫主导抗原集将通过许可协议向商业疫苗开发商使用。多重血清诊断芯片将有助于确定进入这些代理商的地区的军事人员或平民是否暴露于其中任何一个。他们还将受益于公共卫生监测活动，以跟踪流行地区感染流行的年度变化，并监测发展中国家的血液供应。这些芯片也将有助于评估生物恐怖袭击后人群中暴露的传播。