An Improved Diagnostic for Lyme Arthritis

描述（由申请人提供）： 每年，成千上万的美国人感染莱姆病，这是由细菌伯氏疏螺旋体引起的。目前，莱姆病的临床诊断受到低水平B的限制。临床样品中的伯氏细胞。这对那些患有这种疾病的人来说是一个严重的困难，因为大多数未经治疗的人都会患上莱姆关节炎。莱姆关节炎是一种衰弱的条件表现为急性关节疼痛和肿胀的症状。在这项提案中，我们概述了一个研究项目，将开发一个高度敏感的，莱姆病的诊断检测。与目前可用的其他检测试剂盒不同，该检测试剂盒将直接检测B。本发明涉及在患有莱姆关节炎的个体的滑液中的伯氏细胞，从而填补了目前市场上存在的空白。值得注意的是，数千名患有关节炎症状的美国人接受了一系列测试，以帮助确定他们不适的根源。该分析将为这些患者提供有价值的信息，提供对治疗方案的深入了解。 在这个I期计划中，我们开始开发一种敏感的莱姆病关节炎诊断方法。我们将鉴定直接检测B的高亲和力肽配体。burgdorferi细胞通过它们与细菌外表面蛋白ErpA和ErpP的相互作用。本试验的肽序列将来源于人补体调节因子H，其是ErpA和ErpP的已知配体。我们将确定和亲和力成熟的肽序列的因子H特异性结合ErpA和ErpP使用高密度肽芯片选择。B。将进行Burgdorferi全细胞结合测定以检查为我们的诊断测定选择的每种肽的效用。此外，我们将使用人类细胞进行全细胞结合试验，以证明我们的诊断探针的特异性。一旦我们鉴定出特异性检测B的高亲和力肽。通过它们与外表面蛋白ErpA和ErpP的相互作用，在全细胞结合测定中对burgdorferi细胞进行研究，这些配体将进展到我们的莱姆关节炎诊断计划的第二阶段，该计划将包括开发用于诊断莱姆关节炎的原型仪器。 值得注意的是，我们为这项提案组建了一个强大的研究团队。主要研究者John Mueller博士是一位分子微生物学家，在药物发现的肽选择以及体内和体外生物测定的开发方面拥有丰富的经验。Sriram Shankar博士在设计和开发基于抗体和肽的检测方面经验丰富。布赖恩史蒂文森博士是国际知名的B。Burgdorferi微生物学家，他的研究重点是外表面蛋白ErpA和ErpP的生物学及其与人类因子H的相互作用。总之，我们认为，考虑到因子H和B之间的强相互作用。通过对伯氏螺旋体外表面蛋白ErpA和ErpP的研究，我们将成功地鉴定出可以特异性检测B的高亲和力肽。burgdorferi细胞在我们的全细胞结合试验中。这些配体将作为高亲和力探针，在我们的新的莱姆病诊断检测。 公共卫生相关性： 我们建议开发一种新的莱姆病关节炎的诊断方法，将直接检测B。临床样品中的伯氏细胞。我们将鉴定来源于人补体调节因子H的高亲和力肽，其将共同用作特异性结合B的诊断探针。Burgdorferi外表面蛋白ErpA和ErpP。这些肽配体将为我们提供一种简便的检测B的方法。临床样品中的伯氏细胞。

DESCRIPTION (provided by applicant): Every year, thousands of Americans contract Lyme disease, which is caused by the bacterium Borrelia burgdorferi. Currently, clinical diagnosis of Lyme disease is limited by the low levels of B. burgdorferi cells in clinical samples. This presents a severe hardship on those who have the disease, since most individuals who go untreated develop Lyme arthritis. Lyme arthritis is a debilitating condition manifested by symptoms of acute joint pain and swelling. In this proposal, we outline a research project that will develop a highly sensitive, diagnostic assay for Lyme disease. This assay, unlike other assays that are currently available, will directly detect B. burgdorferi cells in the synovial fluid of individuals suffering from Lyme arthritis, thus filling a void that currently exists in the market. Notably, thousands of Americans who suffer with arthritic symptoms undergo a battery of tests to help pinpoint the source of their discomfort. This assay will provide valuable information to those patients, offering insight into treatment options. In this Phase I proposal, we initiate development of a sensitive diagnostic assay for Lyme arthritis. We will identify high affinity peptide ligands that directly detect B. burgdorferi cells through their interaction with the bacterial outer surface proteins ErpA and ErpP. Peptide sequences for this assay will be derived from the human complement regulator, factor H, which is a known ligand of ErpA and ErpP. We will identify and affinity mature peptide sequences of factor H that specifically bind ErpA and ErpP using high-density peptide chip selections. B. burgdorferi whole-cell binding assays will be performed to examine the utility of each of the peptides selected for our diagnostic assay. In addition, we will carry out whole-cell binding assays using human cells to demonstrate the specificity of our diagnostic probes. Once we have identified high affinity peptides that specifically detect B. burgdorferi cells in whole-cell binding assays through their interactions with outer surface proteins ErpA and ErpP, these ligands will progress to the second Phase of our Lyme Diagnostic program, which will include the development of a prototype instrument for the diagnosis of Lyme arthritis. It is worth noting that we have assembled a strong research team for this proposal. Dr. John Mueller, the Principal Investigator, is a molecular microbiologist who has extensive experience in peptide selections for drug discovery and the development of in vivo and in vitro biological assays. Dr. Sriram Shankar is experienced in the design and development of antibody and peptide-based assays. Dr Brian Stevenson is an internationally renowned B. burgdorferi microbiologist, whose research focuses on the biology of outer surface proteins ErpA and ErpP and their interaction with human factor H. In summary, we feel that given the strong interactions between factor H and the B. burgdorferi outer surface proteins ErpA and ErpP, we will successfully identify high-affinity peptides that can specifically detect B. burgdorferi cells in our whole-cell binding assay. These ligands will serve as high affinity probes in our novel diagnostic assay for Lyme disease. PUBLIC HEALTH RELEVANCE: We propose to develop a novel diagnostic assay for Lyme arthritis that will directly detect B. burgdorferi cells in clinical samples. We will identify high affinity peptides derived from human complement regulator, factor H, that will collectively serve as our diagnostic probe to specifically bind B. burgdorferi outer surface proteins ErpA and ErpP. These peptide ligands will afford us a facile assay to detect B. burgdorferi cells in clinical samples.