A New Cytokine-Based Immunoassay for the Diagnosis of Lyme Disease

用于诊断莱姆病的新的基于细胞因子的免疫测定法

• 批准号: 8877395
• 负责人: Linda K. Bockenstedt
• 金额: $ 48.59万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-06-01 至 2017-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8877395
• 关键词: Affect; Aftercare; Antibiotic Therapy; Antibiotics; Antibodies; Antigens; Autoimmune Diseases; B-Lymphocytes; Biological Assay; Bite; Blood specimen; Borrelia burgdorferi; Cell Communication; Clinical; Communicable Diseases; Complement; Detection; Diagnosis; Diagnostic; Disease; Enzyme-Linked Immunosorbent Assay; Epitopes; Exposure to; Geographic Distribution; Gold; Health Expenditures; Heart; Immunoassay; Immunoblotting; Immunoglobulin G; In Vitro; Incidence; Infection; Infectious Skin Diseases; Inflammatory; Ixodes; Joints; Laboratories; Lead; Lyme Disease; Maps; Measures; Monitor; Morbidity - disease rate; Mus; Nervous system structure; Newly Diagnosed; Order Spirochaetales; Organism; Patients; Peptides; Phase; Physicians; Preventive; Production; Prospective Studies; Proteins; Proteomics; Public Health; Recombinants; Resolution; Rheumatism; Sensitivity and Specificity; Serologic tests; Site; Skin; Specificity; Staging; Subgroup; Symptoms; Syndrome; T cell response; T-Lymphocyte; Techniques; Testing; Ticks; Tissues; Treatment Efficacy; United States; Vector-transmitted infectious disease; Whole Blood; accurate diagnosis; base; cytokine; improved; novel; novel diagnostics; prevent; response; synthetic peptide; tick borne spirochete

PROJECT SUMMARY Lyme disease (LD), caused by the Ixodes tick-borne spirochete Borrelia burgdorferi (Bb), is the most common vector-borne disease in the United States. Despite public health preventive measures, the annual confirmed case incidence has risen to nearly 30,000, the vast majority of which occur in the Northeast. Disseminated infection causes disease in the skin, heart, joints and nervous system and can be more difficult to treat. Although antibiotics achieve clinical cure when administered in early stages of infection, delay in diagnosis and treatment can lead to substantial morbidity and health care expenditures for unexplained symptoms that may persist. Timely and accurate diagnosis of LD, therefore, is essential for optimizing treatment and preventing long-term sequelae of the disease. Currently, serologic tests that detect Bb-reactive antibodies are the mainstay of LD diagnostics, but have low sensitivity early in infection, do not distinguish previous exposure to Bb from active infection, and cannot be used to assess response to therapy. This proposal seeks to improve upon current serologic tests by developing a new diagnostic cytokine assay for LD based on a novel panel of Bb antigens expressed in the skin and/ or required for Bb infection and persistence in the mammalian host. The R21 phase of the proposal will first use a multiplex cytokine assay to determine whether recombinant Bb proteins/synthetic peptides elicit cytokine production from whole blood obtained from patients with newly diagnosed, active LD. We will then optimize the assay conditions, including cytokine selection and Bb protein combination, to produce a refined whole blood cytokine assay that will be further validated in the R33 phase. In a prospective study of a large number of LD subjects and controls conducted during the R33 phase, we will define the sensitivity and specificity of the whole blood cytokine assay and determine whether positive responses in the assay are associated with persistent symptoms after antibiotic treatment. As a strategy that complements current serologic tests, the whole blood cytokine assay will improve the combined sensitivity and specificity of LD diagnostics, especially in situations in which the serologic tests alone under-perform.

项目概要 莱姆病 (LD) 由蜱传螺旋体伯氏疏螺旋体 (Bb) 引起，是最常见的疾病 美国的媒介传播疾病。尽管采取了公共卫生预防措施，但每年确诊的 病例发病数已升至近3万例，其中绝大多数发生在东北地区。传播 感染会导致皮肤、心脏、关节和神经系统疾病，并且可能更难以治疗。 虽然抗生素在感染早期使用时可实现临床治愈，但会延误诊断和治疗。 治疗可能会导致大量的发病率和无法解释的症状的医疗保健支出，这些症状可能 坚持。因此，及时、准确地诊断 LD 对于优化治疗和预防至关重要 该病的长期后遗症。目前，检测 Bb 反应性抗体的血清学检测是 LD 诊断的主要手段，但在感染早期敏感性较低，无法区分之前的暴露情况 Bb 来自活动性感染，不能用于评估治疗反应。该提案旨在改进 基于当前的血清学测试，开发了一种新的 LD 诊断细胞因子测定法，该测定法基于一组新的 Bb 抗原在皮肤中表达和/或 Bb 感染和在哺乳动物宿主中持续存在所需。 该提案的R21阶段将首先使用多重细胞因子测定来确定重组Bb是否 蛋白质/合成肽会从新患新冠肺炎患者的全血中引发细胞因子的产生 诊断为活动性 LD。然后我们将优化检测条件，包括细胞因子选择和 Bb 蛋白 组合，以产生精细的全血细胞因子测定，并将在 R33 阶段进一步验证。在 在 R33 阶段对大量 LD 受试者和对照进行的前瞻性研究，我们将 定义全血细胞因子测定的敏感性和特异性并确定是否呈阳性 检测中的反应与抗生素治疗后的持续症状有关。作为一项策略， 作为当前血清学测试的补充，全血细胞因子测定将提高综合敏感性和 LD 诊断的特异性，特别是在单独血清学检测表现不佳的情况下。