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: 8839960
• 负责人: Linda K. Bockenstedt
• 金额: $ 49.17万
• 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/8839960
• 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; 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.

项目摘要 莱姆病（LD）是由硬蜱传播的螺旋体伯氏疏螺旋体（Bb）引起的最常见的疾病 病媒传播的疾病尽管采取了公共卫生预防措施， 发病病例已上升至近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、关节和神经系统的疾病，而且可能更难治疗。 虽然抗生素在感染的早期阶段使用时可以达到临床治愈，但诊断延迟和 治疗可能导致大量的发病率和无法解释的症状的医疗保健支出， 坚持。因此，及时准确地诊断LD对于优化治疗和预防LD的发生至关重要。 疾病的长期后遗症。目前，检测BB反应性抗体的血清学试验是 LD诊断的主流，但在感染早期具有低灵敏度，不能区分先前暴露于 活动性感染，不能用于评估对治疗的反应。这项建议旨在改善 通过开发一种新的诊断LD的细胞因子检测方法， Bb抗原在皮肤中表达和/或在哺乳动物宿主中Bb感染和持久性所需。 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 蛋白质/合成肽引起来自全血的细胞因子产生，所述全血获自患有新 确诊的活动性LD然后我们将优化测定条件，包括细胞因子选择和Bb蛋白 组合，以产生将在R33阶段进一步验证的精制全血细胞因子测定。在 在R33阶段进行的大量LD受试者和对照的前瞻性研究，我们将 确定全血细胞因子测定的灵敏度和特异性， 测定中的反应与抗生素治疗后的持续症状有关。作为一种战略， 补充目前的血清学试验，全血细胞因子测定将提高综合灵敏度， LD诊断的特异性，特别是在单独的血清学测试表现不佳的情况下。