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

描述（由申请人提供）：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。早期感染的治疗非常有效，早期诊断和治疗对于预防疾病进展至关重要。相比之下，弥散性晚期感染与衰弱有关，有时会对神经和肌肉骨骼系统造成永久性损害，并且可能对抗生素无效。目前，莱姆病的实验室诊断是基于在两层血清学检测中检测伯氏疏螺旋体抗体。然而，使用伯氏疏螺旋体的天然或重组蛋白作为抗原的血清学检测通常对早期莱姆病患者通常寻求初始医疗护理时存在的抗体不敏感和/或缺乏足够的特异性，因为天然和重组伯氏疏螺旋体蛋白抗原都含有B细胞表位，可与针对其他细菌种类的抗体交叉反应。结果，它一直是

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. PUBLIC HEALTH RELEVANCE: Lyme disease is a clinically progressive disease that can result in permanent debilitating neurological and musculoskeletal damage if the infection is allowed to persist and become disseminated. Early treatment is effective and is critical for the prevention of disseminated disease; however, the currently available diagnostic serologic tests lack sufficient specificity and sensitivity during early disease, and fail to correctly diagnose Lye disease in patients as often as 50% of the time. This application takes an entirely new approach to the design of Lyme disease diagnostics, by focusing on the detection of disease specific T cells in infected individuals using unique peptide antigens to overcome the problems of sensitivity and specificity that plague protein-based serological assays. The result will be the development of an effective diagnostic assay for Lyme disease that will improve disease outcomes in patients through early detection, allowing treatment prior to dissemination.