Multiplex Serodiagnostic Chip for Rickettsial Diseases

立克次体病多重血清诊断芯片

• 批准号: 7649856
• 负责人: Juan P Olano
• 金额: $ 9.29万
• 依托单位: UNIVERSITY OF TEXAS MED BR GALVESTON
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-03-01 至 2009-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7649856
• 关键词: Agreement; Animals; Antibody Formation; Antigens; Bacteria; Biological Assay; Businesses; Cell-Free System; Communicable Diseases; Guanosine Monophosphate; Human; Immune response; Immunodominant Antigens; Individual; Infectious Agent; Licensing; Methods; Open Reading Frames; Parasites; Patients; Performance; Polymerase Chain Reaction; Printing; Proteins; Proteome; Research Personnel; Rickettsia; Rickettsia Infections; Sampling; Sensitivity and Specificity; Serum; Technology; Testing; Vaccine Antigen; Virus; base; high throughput analysis; improved; microorganism

We have developed technology that enables rapid, compI"ehensive and high-throughput analysis of humoral immune responses to infectious disease antigens that is being used fOf subunit vaccine antigen discovery and to develop serodiagnostic tests. The method is based on printing microarray chips with proteins encoded by infectious microorganisms. Currently, we have expressed and printed more than 16,000 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, and identify a sel 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 confIQured. Here we wiD fabricate whole proteome miaoarrays for Rickettsia prowazekH and R. rickettsii. The arrays '11 be probed with serum from well-<:haracterized infected humans and animals. and healthy controls 10 identify the immunodominant and early reactive serodiagnostic antigen sets. The ability of this chip to distinguish welkharacterized sera collected from humans and animals infected with the different rlc:kettsiae '1 be determined and the results compared with established assays done on the same samples. Specific Aims 1. PCR and done 2246 ORFs from two rickettsiae. 2. Express each ORF in Ihe cell free system and print each protein onto Microarray chips. 3. Probe the microarrays with well characterized human and animal sera to identify the immunodominant antigen set for each agent. 4. Transfer the immunodominant antigen sets to immunostrips and validate the assay perfonnance with well characterized infected sera. The p1asmids, proteins. and protein mictoarrays generated from this project will be available to other RCE investigators ltv"ough the PSWRCE Protein Miaoarray Core. The validated immunostrips that result from this project win be transferred under a licensing agreement to a commercial business for GMP manufactUring.

我们开发了能够快速，兼容的技术 对使用FOF亚基疫苗抗原的传染病抗原的体液免疫反应 发现并开发血清诊断测试。该方法基于印刷微阵列芯片 由传染性微生物编码的蛋白质。目前，我们已经表达并印刷了超过16,000 来自6种细菌，17种病毒和2个寄生虫的单个蛋白质。芯片用血清探测 感染患者和健康对照，以鉴定主要的抗体反应，并确定10-20的SER 每种感染剂的血清诊断抗原。通过对每个感染性使用多种抗原 代理，可以将具有统计学上提高灵敏度和特异性的血清诊断测试构成约束。 在这里，我们为Rickettsia Prowazekh和R. Rickettsii制造了整个蛋白质组的序列。阵列 '11用井中的血清探测 - <：受害的人和动物。和健康对照10 确定免疫主导和早期反应性血清诊断抗原套件。这个芯片的能力 区分被不同RLC感染的人类和动物收集的welkharaCharaCharatized血清：kettsiae '1可以确定，结果与在相同样品上进行的既定测定法相比。 具体目标 1。PCR并从两个立克的2246个ORF完成。 2。在无细胞系统中表达每个ORF，然后将每个蛋白质打印到微阵列芯片上。 3。探测具有良好特征的人类和动物血清的微阵列，以识别免疫力学 为每个代理设置抗原。 4。将免疫主导抗原套件转移到免疫带中，并用孔验证测定法 表征感染的血清。 P1asmid，蛋白质。该项目产生的蛋白质小动物将提供给其他RCE 研究人员LTV“ PSWRCE蛋白MiaoArray核心。 根据许可协议将项目获胜转让给GMP制造商的商业业务。