Cross-species serum antibody detection by direct immunoglobulin catalysis

目前医学上对莱姆病的诊断采用症状观察结合血液检测的方法，存在很多错误。长期目标是开发一种新方法，克服莱姆病检测中的这一限制，并创建一种可用于人类、宠物、牲畜和野生动物的莱姆病通用（跨物种）检测方法。这项拟议的研究利用了莱姆病感染后形成的抗体的一个很大程度上未知的特性，可以通过它们的化学反应性来方便地测量。莱姆病是美国最常见的媒介传播疾病，每年感染30万美国人。目前的医学做法是通过观察症状，结合ELISA和Western blot检测宿主动物抗伯氏疏螺旋体抗原抗体的血清学检测来诊断莱姆病。这两种方法都有其局限性，即二级试剂表现出非特异性结合。二抗使测试对提供血清的物种具有特异性。廉价、同型和物种无关的抗体传感器很重要，因为它们可以促进传染病的广泛监测、血清转化动力学的跨物种研究和接触后的早期检测。长期目标是实现和标准化一种新的基于抗体催化水氧化途径（ACWOP）直接测量抗体催化的生物传感器。目的是阐明抗原固定的物理化学和催化传感产生过氧化氢的比色或电化学读数。将追求三个研究目标；即通过实验证明抗原定位和特异性抗体在纸底物上的结合；结合抗体的直接催化读数显示elisa水平的敏感性；并且，在临床相关样本中检测到伯氏疏螺旋体抗原。关键特点是新的acwop转导机制，消除了免疫特异性传感器的二抗，以及对非特异性结合具有抗性的固定化学的实际实施。预计所提出的生物传感器将具有重大的实际影响，因为莱姆病的非物种检测可以监测博氏螺旋体水库以及牲畜，伴侣动物和人类。

Current medical practice to diagnose Lyme disease by observation of symptoms combined with blood testing produces a lot of errors. The long-term goal is to develop a new approach that overcomes this limitation in Lyme disease testing and to create a general (cross­species) test for Lyme disease that can be used in humans, pets, livestock, and wild animals. The proposed research takes advantage of a largely-unknown property of the antibodies that form in response to Lyme infection that can be conveniently measured by their chemical reactivity. Lyme disease is the most common vector­borne disease in the United States, infecting 300,000 Americans per year. Current medical practice is to diagnose Lyme disease by observation of symptoms combined with serological detection of the host animal's antibodies against Borrelia antigens using ELISA and Western blot. Both of them have their limitations, namely, the secondary reagents exhibit nonspecific binding. The secondary antibodies make the test specific to the species providing the serum. Inexpensive, isotype- and species-independent antibody sensors are important because they can facilitate widespread monitoring of infectious disease, the cross-species study of the dynamics of seroconversion and early detection following exposure. The long-term goal is to implement and standardize a new biosensor based on direct measurement of antibody catalysis via the antibody-catalyzed water oxidation pathway (ACWOP). The objective is to elucidate the physiochemistry of antigen immobilization and catalytic sensing which produces hydrogen peroxide for a colorimetric or electrochemical readout. Three research aims will be pursued; namely, experimentally show antigen localization and specific antibody binding on paper substrates; show ELISA-level sensitivity from direct catalytic readout of bound antibodies; and, show detection of Borrelia antigens in clinically relevant samples. The key features are novel ACWOP-transduction mechanism that eliminates secondary antibodies from immune-specific sensors and practical implementation of immobilization chemistry that is resistant to nonspecific binding. It is anticipated that the proposed biosensor will have a significant practical impact because species­independent detection of Lyme borreliosis enables monitoring of the Borrelia reservoir as well as livestock, companion animals, and humans.