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了莱姆病，这是由细菌伯氏菌burgdorferi引起的。当前，莱姆病的临床诊断受临床样品中伯格多菲利细胞的低水平限制。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会出现莱姆关节炎。莱姆关节炎是一种令人衰弱的疾病，表现为急性关节疼痛和肿胀的症状。在此提案中，我们概述了一个研究项目，该项目将开发出对莱姆病的高度敏感，诊断测定。与当前可用的其他测定法不同，该测定法将直接检测出患有莱姆关节炎的个体的滑动液中的B. burgdorferi细胞，从而填充了当前在市场上存在的空白。值得注意的是，成千上万患有关节炎症状的美国人经历了一系列测试，以帮助查明自己的不适感。该测定法将为那些患者提供有价值的信息，从而洞悉治疗选择。在这一阶段的提案中，我们启动开发莱姆关节炎敏感的诊断测定法。我们将通过与细菌外表面蛋白ERPA和ERPP的相互作用来鉴定高亲和肽配体，这些肽配体可以直接检测B. burgdorferi细胞。该测定法的肽序列将来自人类补体调节剂因子H，这是已知的ERPA和ERPP的配体。我们将识别和亲和力成熟的因子H的成熟肽序列，该因子H使用高密度肽芯片选择特异性结合ERPA和ERPP。 B. burgdorferi全细胞结合测定法将进行检查，以检查选择用于诊断测定的每种肽的效用。此外，我们将使用人类细胞进行全细胞结合测定，以证明我们的诊断探针的特异性。一旦我们确定了高亲和力肽，这些肽通过与外表面蛋白ERPA和ERPP的相互作用来特异性检测全细胞结合测定中的B. burgdorferi细胞，这些配体将发展为我们莱姆诊断程序的第二阶段，其中包括用于诊断莱姆属lyme Arththrthrathrythis的原型仪器的开发。值得注意的是，我们已经为该提案组成了一个强大的研究团队。首席研究员约翰·穆勒（John Mueller）博士是一位分子微生物学家，在药物发现和体内和体外生物学测定方面的肽选择方面拥有丰富的经验。 Sriram Shankar博士在抗体和基于肽的测定的设计和开发方面经验丰富。 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在我们的全细胞结合测定中。这些配体将在我们对莱姆病的新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的莱姆关节炎诊断测定法，该测定法将直接检测到临床样品中的伯氏芽孢杆菌细胞。我们将确定从人体补体调节剂H H的高亲和力肽H，该肽h将共同用作我们的诊断探针，以特异性结合B. burgdorferi外表面蛋白ERPA和ERPP。这些肽配体将为我们提供一个轻松的测定法，以检测临床样品中的爆发芽孢杆菌细胞。