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: 8466282
• 负责人: Linda K. Bockenstedt
• 金额: $ 20.32万
• 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 2014-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8466282
• 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; 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

DESCRIPTION (provided by applicant): 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 infectio, 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.

描述（由申请人提供）：由ixodes tick-borne spirochete borlelia burgdorferi（BB）引起的莱姆病（LD）是美国最常见的媒介传播疾病。尽管采取了公共卫生预防措施，但年度确认的病例发病率已上升到近30,000，其中绝大多数发生在东北。传播感染会导致皮肤，心脏，关节和神经系统的疾病，并且可能更难治疗。尽管抗生素在感染的早期阶段施用时可实现临床治愈 诊断和治疗可能导致可能持续存在的无法解释的症状的大量发病率和医疗保健支出。因此，及时，准确的LD诊断对于优化治疗和预防疾病的长期后遗症至关重要。当前，检测BB反应性抗体的血清学测试是LD诊断的主要主体，但在感染的早期具有较低的灵敏度，不能区分先前接触BB与主动感染的接触，并且不能用于评估对治疗的反应。该建议旨在通过基于在皮肤中表达的新型BB抗原和/或BB感染和哺乳动物宿主持续性所需的新型BB抗原面板开发新的LD诊断细胞因子测定，以改善当前的血清学测试。该提案的R21阶段将首先使用多重细胞因子测定法来确定重组BB蛋白/合成肽是否从新诊断的活性LD患者获得的全血会引起细胞因子产生。然后，我们将优化包括细胞因子选择和BB蛋白组合在内的测定条件，以产生精致的全血细胞因子测定法，该测定将在R33期间进一步验证。在对R33阶段进行的大量LD受试者和对照的前瞻性研究中，我们将定义整个血细胞因子测定的敏感性和特异性，并确定测定中的阳性反应是否与抗生素治疗后的持续症状有关。作为对当前血清学测试的补充的策略，整个血细胞因子测定法将提高LD诊断的综合敏感性和特异性，尤其是在单独使用血清学测试的情况下。