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: 8042576
• 负责人: Joseph Frank Petrosino
• 金额: $ 44.33万
• 依托单位: UNIVERSITY OF TEXAS MED BR GALVESTON
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8042576
• 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

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.

检测生物防御病原体的诊断试剂和检测方法是公共安全的关键需求。两把钥匙 成功的分子诊断测定的组件是直接与 目标生物体，或生物体分泌的产物，以及指示的信号域，具有很大的 当传感器结合目标分子时的灵敏度。在过去的 18 个月里，我们实施了 成熟的技术：噬菌体展示、共价蛋白-DNA 连接、实时 PCR 以及结果 从我们的抗原发现研究开始，开始创建一种强大的诊断方法来检测是否存在 生物和环境样本中的土拉弗朗西斯菌 (Ft) 以及针对的免疫反应 反对英尺。这些检测的核心蛋白质-DNA 嵌合体称为蝌蚪，它们能够 与类似的酶联技术相比，灵敏度更高（约 106 倍） 免疫吸附测定（ELISA）。此外，这些诊断能够识别目标分子 浓度动态范围宽。 在本提案中，我们将多重蝌蚪检测，包括额外的发烧和腹泻药物，如 总体 WRCE 诊断主题计划中概述。 Ft、裂谷热病毒和小隐孢子虫 将作为平台 WRCE 诊断方法和代理列表的初始比较控制 根据WRCE计划，将在未来几年进行扩展。此外，我们将寻求整合 WRCE 诊断小组正在开发的其他平台的基本技术，例如横向 流动微流体，扩大蝌蚪诊断的实用性，因为它们被进一步复用于 同时检测多种试剂，并且除其用途外还适用于护理点使用 在参考实验室。