Multi-peptide assay for serodiagnosis of Lyme disease

描述（由申请人提供）：我们建议开发一种更灵敏、更特异的血清学检测方法，用于检测莱姆病，尤其是早期疾病。莱姆病是美国和欧洲最常见的病媒传染病，因此是一个重要的公共卫生问题。该疾病影响多个器官系统，包括肌肉骨骼、皮肤和神经系统，并包括在多种疾病的鉴别诊断中。在没有游走性红斑（早期莱姆病的经典皮肤病变）的情况下，通过检测具有提示该疾病的客观发现的患者中对伯氏疏螺旋体的抗体应答来建立诊断。及时诊断很重要，因为莱姆病的早期治疗限制或防止了对受影响系统的严重损害。目前的血清学诊断缺乏早期疾病的敏感性。有几个B。在人类感染早期识别的伯氏螺旋体抗原如FlaB、OspC、VlsE、BBK 32和DbpA。当单独用于血清学测定时，这些抗原中没有一种是足够的。然而，结合起来，它们可以为大大改进的测定提供基础。线性表位已在关键B中鉴定。Burgdorferi抗原如VlsE的恒定区6（IR 6），证明含有这些表位的肽可用作诊断靶。在本I期STTR中，我们建议分析FlaB、几组OspC、BBK 32和DbpA，以定义其他抗原性线性表位。然后，我们将合成肽，并比较它们与相应重组蛋白的血清学反应性。发现的新的免疫诊断肽将被添加到改进的VlsE-IR 6肽中，以开发新的、更灵敏的免疫诊断测定。在随后的第二阶段提案中，我们计划筛选和优化免疫诊断肽的组合，并准备在ELISA和侧流格式中验证新的免疫测定。最终，我们的目标是用新一代的基于多肽的测试取代今天的检测。由于我们可以实现更高的特异性和灵敏度的提高，这种方法可能导致开发一种单层检测方法来检测针对B的三种致病基因种中任何一种的抗体。莱姆病的早期和晚期都有burgdorferi。 莱姆病是美国最常见的媒介传染病，影响多个器官系统。虽然及时的诊断和治疗可以防止或限制对受影响系统的严重损伤，但目前莱姆病的血清诊断在早期疾病中缺乏敏感性，并且不足以单独使用。我们的目标是用一种新的多肽测试取代目前的检测方法。由于我们可以实现更高的特异性和灵敏度的提高，这种方法可能导致开发一种单层检测方法来检测针对B的三种致病基因种中任何一种的抗体。莱姆病的早期和晚期都有burgdorferi。

DESCRIPTION (provided by applicant): We propose to develop a more sensitive, specific serologic assay for detection of Lyme disease especially early disease. Lyme disease is the most common vector borne infectious disease in the United States and Europe and as such it is a significant public health concern. The disease affects multiple organ systems including musculoskeletal, skin and nervous system and is included in the differential diagnosis of multiple diseases. In the absence of erythema migrans, the classic skin lesion of early Lyme disease, the diagnosis is established by the detection of an antibody response to Borrelia burgdorferi in patients with objective findings suggestive of the disease. Prompt diagnosis is important because early treatment of Lyme disease limits or prevents serious damage to the systems affected. Current serodiagnostics lack sensitivity in early disease. There are a number of B. burgdorferi antigens recognized early in human infection such as FlaB, OspC, VlsE, BBK32 and DbpA. None of these antigens is adequate when used alone in serologic assays. However, in combination they could provide the basis for a much improved assay. Linear epitopes have been identified in key B. burgdorferi antigens such as the invariable region 6 (IR6) of VlsE, proving that peptides containing these epitopes can be used as diagnostic targets. In this Phase I STTR, we propose to analyze FlaB, several groups of OspC, BBK32 and DbpA to define additional antigenic linear epitopes. We will then make synthetic peptides and compare their serologic reactivity to the corresponding recombinant protein. The new immunodiagnostic peptides discovered will be added to an improved VlsE-IR6 peptide to develop new, more sensitive immunodiagnostic assays. In a subsequent Phase II proposal we plan to screen and optimize a combination of immunodiagnostic peptides and prepare for validation of new immunoassays in both ELISA and lateral flow formats. Ultimately, our goal is to replace today's assays with a new generation of multi-peptide based tests. Because of greater specificity and the improvement of sensitivity that we can achieve, this approach could lead to the development of a single tier assay to detect antibodies against any of the three pathogenic genospecies of B. burgdorferi in both early and late Lyme disease. Lyme disease, the most common vector borne infectious disease in the United States affects multiple organ systems. Although prompt diagnosis and treatment prevents or limits serious injury to the systems affected, current serodiagnostics for Lyme disease lack sensitivity in early disease and are not specific enough to be used alone. Our objective is to replace today's assays with a new multi-peptide test. Because of greater specificity and the improvement of sensitivity that we can achieve, this approach could lead to the development of a single tier assay to detect antibodies against any of the three pathogenic genospecies of B. burgdorferi in both early and late Lyme disease.