Approach for amplification of molecular signals

分子信号放大方法

• 批准号: 6935526
• 负责人: Alexey G Zdanovsky
• 金额: $ 16.47万
• 依托单位: ALPHA UNIVERSE, LLC
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-01 至 2007-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6935526
• 关键词: DNA directed DNA polymerase; Escherichia coli; antimicroorganism antibody; bacterial proteins; bacteriophage lambda; binding proteins; biohazard detection; bioluminescence; biotechnology; bioterrorism /chemical warfare; biotin; capsid; enzyme linked immunosorbent assay; immunologic assay /test; luciferin monooxygenase; nucleic acid amplification techniques; plasmids; recombinant virus; reporter genes; technology /technique development; transfection /expression vector; virus genetics

DESCRIPTION (provided by applicant): Antibody-based detection systems have been used for years in clinical diagnostics and environmental monitoring for the detection and identification of pathogenic agents. Recent advances in the field of low-level antigen detection utilize the DNA polymerase-mediated signal amplification approach that provides a substantial enhancement in detection sensitivity over the conventional ELISA. However, protocols utilizing this approach have not been widely used in diagnostic laboratories because they require rigorous sample preparation, complex reactive components of limited shelf life, expensive hardware and involve a complex detection process. The goal of the current application is to develop an alternative approach for the signal amplification. Bacterial luciferase will be incorporated in the naturally existing cascades of enzymatic reactions to produce an inexpensive and simple assay for the detection of antigens of interest including those of biowarfare agents.

描述（由申请人提供）：基于抗体的检测系统已在临床诊断和环境监测中使用多年，用于检测和鉴定病原体。低水平抗原检测领域的最新进展利用DNA聚合酶介导的信号放大方法，该方法提供了比常规ELISA显著增强的检测灵敏度。然而，利用这种方法的方案尚未在诊断实验室中广泛使用，因为它们需要严格的样品制备、有限保质期的复杂反应组分、昂贵的硬件并且涉及复杂的检测过程。本申请的目标是开发用于信号放大的替代方法。细菌荧光素酶将被并入酶促反应的天然存在的级联中，以产生用于检测感兴趣的抗原（包括生物战剂的抗原）的廉价且简单的测定。