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

基本信息

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
项目类别：
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

项目摘要

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）：莱姆病（LD）由蜱传螺旋体伯氏疏螺旋体（Bb）引起，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，每年确诊病例数仍上升至近3万例，其中绝大多数发生在东北地区。播散性感染会导致皮肤、心脏、关节和神经系统疾病，并且更难以治疗。尽管在感染早期阶段使用抗生素可以实现临床治愈，但延迟了诊断和治疗可能会导致大量的发病率和可能持续存在的不明原因症状的医疗费用。因此，及时准确地诊断 LD 对于优化治疗和预防该疾病的长期后遗症至关重要。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，无法区分先前接触过 Bb 的患者与活动性感染，并且不能用于评估对治疗的反应。该提案旨在通过开发一种新的 LD 诊断细胞因子测定法来改进当前的血清学测试，该测定法基于皮肤中表达的和/或 Bb 感染和在哺乳动物宿主中持续存在所需的一组新的 Bb 抗原。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/合成肽是否会引发从新诊断的活动性 LD 患者获得的全血中产生细胞因子。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检测，并将在 R33 阶段进一步验证。在 R33 阶段对大量 LD 受试者和对照进行的前瞻性研究中，我们将定义全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相关。作为补充当前血清学检测的策略，全血细胞因子测定将提高 LD 诊断的综合敏感性和特异性，特别是在单独血清学检测表现不佳的情况下。