SBIR Phase II: Inexpensive and Effecient System for Signal Amplification

该小型企业创新研究（SBIR）第二阶段项目提出开发一种新的信号放大系统，该系统价格低廉且使用简单，可与抗体或其他探针分子结合，并可将常规酶联免疫吸附测定（ELISA）的检测限提高100至1000倍。ELISA已在科学和诊断中使用了40多年。ELISA是一种简便、可靠、廉价的方法。然而，由于在该技术中使用的酶作为信号产生和放大装置的性质，其具有相对低的灵敏度。在阶段I期间，证明了用可以传播并同时产生光信号的β-内酰胺酶取代这些酶显著增加了ELISA的灵敏度，同时降低了测定的成本，而不会使其方案复杂化。第二阶段的目标是进一步提高系统的灵敏度和响应时间，并创造两种类型的产品：1）可插入常规ELISA系统的组件，以及2）用于检测特定毒素和其他病原体的试剂盒。该项目的更广泛的影响/商业潜力预计将影响基础研究，生物防御和医学领域。所提出的系统将允许非常灵敏和廉价的抗原检测的选择。对于那些参与基础研究的人来说，拟议的免疫测定将允许对生物过程进行更详细的分析，甚至可能为监测以前无法检测到的过程和分子打开大门。在生物防御和医学诊断方面，提高灵敏度意味着通过允许更早地检测有害病原体以及人类有机体的病理变化来提高安全性。

This Small Business Innovation Research (SBIR) Phase II project proposes to develop a new signal amplification system that, while being inexpensive and simple to use, can be combined with antibodies or other probe molecules and can increase detection limits of conventional Enzyme-linked immunosorbent assay (ELISA) by 100 to 1000-fold. ELISA has been used both in science and diagnostic for more than 40 years. ELISA is simple, reliable, and inexpensive technique. However, because of the nature of enzymes used in this technique as signal-generating and amplifying device, it has a relatively low sensitivity. During Phase I, it was demonstrated that substitution of these enzymes with phages that can propagate and simultaneously generate light signal substantially increases the sensitivity of ELISA and simultaneously reduces cost of the assay without complicating its protocol. In Phase II, the goal is to further improve sensitivity and response time of the system and create two types of products: 1) components that can be plugged-in into conventional ELISA system, and 2) kits designed for detection of specific toxins and other pathogens.The broader impact /commercial potential of this project is expected to impact the areas of basic research, biodefense, and medicine. The proposed system will allow very sensitive and inexpensive detection of antigens of choice. For those involved in basic research, the proposed immunoassay will allow a significantly more detailed analysis of biological processes, possibly even opening the doors to monitoring those processes and molecules previously undetectable. In biodefense and medical diagnostics, increased sensitivity will mean an increased security by allowing earlier detection of harmful pathogens, as well as pathological changes in the human organism.