Development of a Cytokine Release Assay as a Diagnostic Test for Lyme Disease

描述(申请人提供)：莱姆病是北美和欧洲最常见的媒介传播传染病。它是一种进行性疾病，具有广泛的临床表现。早期感染的治疗是非常有效的，早期诊断和治疗是防止疾病进展的关键。相比之下，播散性晚期感染会对神经和肌肉骨骼系统造成衰弱的、有时是永久性的损害，而且对抗生素可能是无效的。目前莱姆病的实验室诊断是基于两级血清学检测伯氏疏螺旋体抗体。然而，使用伯氏杆菌天然或重组蛋白作为抗原的血清学检测往往对检测存在的抗体不敏感，因为许多早期莱姆病患者通常寻求初步医疗和/或缺乏足够的特异性，因为天然和重组伯氏杆菌蛋白抗原都包含与其他细菌物种的抗体交叉反应的B细胞表位。因此，它一直是 据估计，目前的IgM或IgG莱姆病检测方法在约50%的时间内无法诊断患者的早期疾病。在疑似莱姆病的患者中，血清学检测未能一致地识别伯氏杆菌感染，因此有必要开发一种新的诊断测试。Quantiferon(R)试验是一种基于细胞的IFNG释放试验，用于检测感染结核分枝杆菌患者血液中的抗原特异性T细胞。这项检测促炎细胞因子干扰素产生的方法是什么？在结核分枝杆菌多肽抗原的刺激下，已被证明在诊断结核病方面具有高度的敏感性和高度的特异性，取代了无效的血清学检测，并改进了使用结核菌素皮肤试验的诊断。基于细胞的检测将代表莱姆病诊断检测方法发展的新方向，有可能比目前的血清学检测方法更具特异性和敏感性。然而，这种分析的发展需要鉴定伯氏杆菌特有的高度特异的多肽表位。目前这项研究的目标是识别来自伯氏杆菌蛋白的独特T细胞表位，用于基于Quantiferon(注册商标)的莱姆病诊断细胞检测。

DESCRIPTION (provided by applicant): Lyme disease is the most common vector borne infectious disease in North America and Europe. It is a progressive disease with a wide array of clinical manifestations. Treatment of early infection is highly effective, and early diagnosis and treatment are critical to prevent disease progression. By contrast, disseminated late- phase infection is associated with debilitating, sometimes, permanent damage to the nervous and musculoskeletal systems and can be refractory to antibiotics. Currently the laboratory diagnosis of Lyme disease is based on the detection of antibodies against Borrelia burgdorferi in a two-tier serological assay. However, serological assays using native or recombinant proteins from B. burgdorferi as antigens are often insensitive for the detection of antibody present at the time many patients with early Lyme disease usually seek initial medical care and/or lack sufficient specificity as both native and recombinant B. burgdorferi protein antigens contain B cell epitopes that cross react with antibodies against other bacterial species. As a result, it has been estimated that current IgM or IgG Lyme disease assays fail to diagnose early disease in patients ~50% of the time. The failure of serological assays to consistently identify B. burgdorferi infection in patients with suspected Lyme disease necessitates the development of a new class of diagnostic tests. The QuantiFERON(R) assay is a cellular-based IFNg release assay for the detection of antigen-specific T cells in the blood of patients infected with Mycobacterium tuberculosis. This assay, which detects the production of the proinflammatory cytokine IFN? in response to stimulation with peptide antigens derived from M. tuberculosis, has proven to be highly sensitive and highly specific in the diagnosis of tuberculosis, supplanting ineffective serological assays and improving upon diagnosis using the tuberculin skin test. A cellular based assay would represent a new direction in the development of diagnostic assays for Lyme disease, with the potential for being significantly more specific and sensitive than current serological assays. However, the development of such an assay requires the identification of highly-specific peptide epitopes unique to B. burgdorferi. The goal of the current study is to identify unique T cell-epitopes derived from B. burgdorferi proteins for use in a QuantiFERON(R)-based cellular assay for the diagnosis of Lyme disease.