An Improved Diagnostic for Lyme Arthritis

莱姆关节炎的改进诊断

基本信息

批准号：
7999233
负责人：
JOHN E MUELLER
金额：
$ 15.03万
依托单位：
LYNNTECH, INC.
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-06-15 至 2011-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7999233
关键词：
Acute Affinity American Amino Acid Sequence Arthralgia Arthritis Bacteria Binding Biological Assay Biology Borrelia burgdorferi Cells Chemicals Clinical Commit Complement Complement Factor H Complex Contracts Detection Development Diagnosis Diagnostic Disease Disease Progression Evaluation Exhibits Filtration Fluorescence Spectroscopy Funding Government Human Human Factor H In Vitro Incidence Individual Label Ligand Binding Ligands Lyme Arthritis Lyme Disease Marketing Medical Membrane Proteins Molecular Mutagenesis Natural Immunity Pathway interactions Patients Peptide antibodies Peptides Persons Phase Preparation Principal Investigator Proteins Relative (related person)Research Research Project Grants Sampling Screening procedure Signal Transduction Source Specificity Swelling Symptoms Synovial Fluid Technology Testing United States authority base clinical Diagnosis commercialization density design drug discovery experience fluorophore improved in vivo innovation insight instrument instrumentation nanoparticle novel diagnostics peptide B point of care programs protein aminoacid sequence prototype public health relevance research and development success tool user-friendly

项目摘要

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.

描述（由申请人提供）：每年，成千上万的美国人感染莱姆病，这是由伯氏疏螺旋体细菌引起的。目前，莱姆病的临床诊断受到临床样本中伯氏疏螺旋体细胞水平低的限制。这给患有这种疾病的人带来了严重的困难，因为大多数未经治疗的人都会患上莱姆关节炎。莱姆关节炎是一种使人衰弱的疾病，表现为急性关节疼痛和肿胀症状。在本提案中，我们概述了一个研究项目，该项目将开发一种高度敏感的莱姆病诊断方法。与目前可用的其他检测不同，该检测将直接检测莱姆关节炎患者滑液中的伯氏疏螺旋体细胞，从而填补了目前市场上存在的空白。值得注意的是，成千上万患有关节炎症状的美国人接受了一系列测试，以帮助查明他们不适的根源。该检测将为这些患者提供有价值的信息，深入了解治疗方案。在这一第一阶段提案中，我们开始开发莱姆关节炎的敏感诊断测定法。我们将鉴定高亲和力肽配体，通过其与细菌外表面蛋白 ErpA 和 ErpP 的相互作用来直接检测伯氏疏螺旋体细胞。用于该测定的肽序列将源自人类补体调节因子 H，它是 ErpA 和 ErpP 的已知配体。我们将使用高密度肽芯片选择来鉴定和亲和特异性结合 ErpA 和 ErpP 的 H 因子成熟肽序列。将进行伯氏疏螺旋体全细胞结合测定，以检查为我们的诊断测定选择的每种肽的效用。此外，我们将使用人体细胞进行全细胞结合测定，以证明我们的诊断探针的特异性。一旦我们通过与外表面蛋白 ErpA 和 ErpP 的相互作用鉴定出在全细胞结合测定中特异性检测伯氏疏螺旋体细胞的高亲和力肽，这些配体将进入我们莱姆病诊断计划的第二阶段，其中包括开发用于诊断莱姆关节炎的原型仪器。值得注意的是，我们为这个提案组建了一个强大的研究团队。首席研究员 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 博士在抗体和肽检测的设计和开发方面经验丰富。 Brian Stevenson 博士是国际知名的伯氏疏螺旋体微生物学家，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，我们认为鉴于因子 H 和 B 之间的强烈相互作用。伯氏疏螺旋体外表面蛋白 ErpA 和 ErpP，我们将在全细胞结合测定中成功鉴定出可以特异性检测伯氏疏螺旋体细胞的高亲和力肽。这些配体将在我们针对莱姆病的新型诊断测定中充当高亲和力探针。公共卫生相关性：我们建议开发一种新型莱姆关节炎诊断方法，可直接检测临床样本中的伯氏疏螺旋体细胞。我们将鉴定源自人补体调节因子 H 的高亲和力肽，它们将共同作为我们的诊断探针，特异性结合伯氏疏螺旋体外表面蛋白 ErpA 和 ErpP。这些肽配体将为我们提供一种简便的检测方法来检测临床样品中的伯氏疏螺旋体细胞。