The Tadpole Assay Multuplex Platform for Biodefense Agent Diagnostics

用于生物防御剂诊断的蝌蚪检测多重平台

• 批准号: 8377045
• 负责人: Joseph Frank Petrosino
• 金额: $ 40.96万
• 依托单位: UNIVERSITY OF TEXAS MED BR GALVESTON
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-03-01 至 2014-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8377045
• 关键词: Address; Affinity; Antibodies; Antigens; Bacteriophage M13; Binding; Biological; Biological Assay; Biological Products; Caring; Cells; Chimera organism; Communicable Diseases; Complex; Cryptosporidium; Cryptosporidium parvum; DNA; Detection; Diagnostic; Diagnostic Reagent; Disease; Elements; Enzyme-Linked Immunosorbent Assay; Epitopes; Fever; Francisella tularensis; Goals; Human; Immune response; Immunodominant Antigens; Immunoglobulin Fragments; Infection; Laboratories; Lateral; Libraries; Ligand Binding; Ligands; Membrane Proteins; Microfluidics; Molecular; Mutagenesis; Nucleic Acids; Organism; Persons; Phage Display; Proteins; Public Health; Reagent; Research; Rift Valley fever virus; Safety; Sampling; Signal Transduction; Site; Surface; Surface Plasmon Resonance; System; Tadpoles; Tail; Technology; Time; Western Blotting; assay development; biodefense; comparative; design; in vivo; member; pathogen; point of care; point-of-care diagnostics; sensor; sugar; Address; Affinity; Antibodies; Antigens; Bacteriophage M13; Binding; Biological; Biological Assay; Biological Products; Caring; Cells; Chimera organism; Communicable Diseases; Complex; Cryptosporidium; Cryptosporidium parvum; DNA; Detection; Diagnostic; Diagnostic Reagent; Disease; Elements; Enzyme-Linked Immunosorbent Assay; Epitopes; Fever; Francisella tularensis; Goals; Human; Immune response; Immunodominant Antigens; Immunoglobulin Fragments; Infection; Laboratories; Lateral; Libraries; Ligand Binding; Ligands; Membrane Proteins; Microfluidics; Molecular; Mutagenesis; Nucleic Acids; Organism; Persons; Phage Display; Proteins; Public Health; Reagent; Research; Rift Valley fever virus; Safety; Sampling; Signal Transduction; Site; Surface; Surface Plasmon Resonance; System; Tadpoles; Tail; Technology; Time; Western Blotting; assay development; biodefense; comparative; design; in vivo; member; pathogen; point of care; point-of-care diagnostics; sensor; sugar

Diagnostic reagents and assays to detect biodefense pathogens are critical needs for public safety. Two key components for a successful molecular diagnostic assay are a sensor component that binds directly to the targeted organism, or to a product secreted by the organism, and a signal domain that indicates, with great sensitivity, when the sensor has bound the target molecule. Over the past 18 months we have implemented proven technologies: phage-display, covalent protein-DNA linkage, and real-time PCR, along with the results from our antigen discovery research, to begin to create a powerful diagnostic assay to detect the presence of Francisella tularensis (Ft) in biological and environmental samples as well as immune responses directed against Ft. The protein-DNA chimeras central to these assays are called tadpoles, which are capable of achieving a much greater level of sensitivity (~106-fold greater) compared to analogous enzyme-linked immunosorbent assays (ELISAs). Furthermore these diagnostics are able to identify target molecules over a wide dynamic range of concentrations. In this proposal, we will multiplex the tadpole assay to include additional fever and diarrheal agents as outlined in the overall WRCE diagnostic theme plan. Ft, Rift Valley fever virus, and Cryptosporidium parvum will serve as initial, comparative controls for the platform WRCE diagnostic approaches, and the agent list will be expanded in later years according to the WRCE plan. Furthermore, we will seek to integrate elemental technologies from other platforms being developed in the WRCE diagnostic group, such as lateral flow microfluidics, to expand the utility of tadpole diagnostics as they are further multiplexed for the simultaneous detection of numerous agents and are adapted for point-of-care usage in addition to their use in the reference laboratory.

诊断试剂和检测生物防腐病原体的测定是公共安全的关键需求。两个键 成功的分子诊断测定的组件是直接结合的传感器成分 有针对性的有机体或有机体分泌的产品，以及指示很大的信号域 当传感器结合靶分子时，灵敏度。在过去的18个月中，我们实施了 经过验证的技术：噬菌体播放，共价蛋白-DNA链接和实时PCR以及结果 从我们的抗原发现研究，开始创建强大的诊断测定法以检测存在 生物和环境样品中的francisella tularensis（ft）以及定向的免疫反应 反对ft。这些测定中心的蛋白质-DNA嵌合体称为t骨，能够 与类似的酶连接相比，达到更高的灵敏度（〜106倍） 免疫吸附测定法（ELISA）。此外，这些诊断能够鉴定在 浓度的动态范围广泛。 在此提案中，我们将在t t t t t thepole分析中多用，以包括额外的发烧和腹泻剂 在整个WRCE诊断主题计划中概述。英尺，裂谷发烧病毒和隐孢子虫 将作为平台WRCE诊断方法的初始比较控制，而代理列表 根据WRCE计划，将在后来的几年进行扩展。此外，我们将寻求整合 WRCE诊断组中开发的其他平台的元素技术，例如横向 流动微流体，以扩大t象诊断的实用性，因为它们被进一步多样地 同时检测众多代理商，并适用于保健点的使用。 在参考实验室。