Multi-antigen peptide assay for the serodiagnosis of Lyme disease

描述(申请人提供)：莱姆病(LD)是美国最常见的媒介传播传染病，仍然是一个重大的公共卫生问题。实验室诊断依赖于伯氏疏螺旋体抗体的检测。目前的血清学检测在早期LD中不够特异，也不敏感。需要新的分析方法，我们计划填补这一未得到满足的需求。我们的主要目标是为LD的血清学诊断提供新的灵敏和特异的多肽分析方法。在第一阶段，我们在确定新的血清学分析的成分方面取得了极大的进展：我们开发了IR6肽，具有更广泛的能力来检测表达不同VlsE序列的螺旋体感染患者的抗体反应性；在17个已知的美国OSPC组中，我们确定了4个(B、F、I和K)作为潜在的表位映射目标；我们评估了由OspC(10个残基)、FLAB(13个残基)和VlsE-IR6(17个残基)序列组成的多抗原肽与LD结合个体抗体的能力。在这个第二阶段的方案中，我们将进一步优化包含OspC、FLAB和IR6表位的多肽，并将验证新的免疫分析方法，包括ELISA法和快速横向流动点(POC)格式。在疾病的早期阶段检测伯氏杆菌抗体具有更高的特异性和更高的灵敏度，可能会导致开发一种能够取代目前推荐的双层范式的单层检测方法。更重要的是，鉴于LD的早期诊断和治疗可以防止疾病的进展和后遗症，这项测试的发展将对改善人类健康做出重要贡献。 公共卫生相关性：莱姆病是美国最常见的媒介传播传染病，影响多个器官系统。尽管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害，但目前的莱姆病血清诊断方法对早期疾病缺乏敏感性，而且不够特异，不能单独使用。我们的目标是用一种新的多抗原肽测试取代目前的检测方法。由于这种方法具有更高的特异性和更高的灵敏度，因此可以开发一种单层检测方法来检测针对三种致病基因种OFB的抗体。莱姆病早期和晚期均有伯氏杆菌感染。

DESCRIPTION (provided by applicant): Lyme Disease (LD) is the most common vector-borne infectious disease in the United States and remains a significant public health concern. The laboratory diagnosis of LD depends on the demonstration of antibodies against Borrelia burgdorferi. Current serologic assays are not specific enough and not sensitive in early LD. New assays are needed and we plan to fill this unmet need. Our main objective is to produce new sensitive and specific peptide based assays for the serodiagnosis of LD. In Phase I we made excellent progress in defining components for a new serologic assay: we developed IR6 peptides with a broader ability to detect antibody reactivity among patients infected with spirochetes expressing different VlsE sequences; of the 17 known US OspC groups we have identified four (B, F, I, and K) as potential targets for epitope mapping; and we evaluated the ability of a multi-antigenic peptide comprised of sequences from OspC (10 residues), FlaB (13 residues) and VlsE-IR6 (17 residues) to bind antibody from individuals with LD. In this Phase II proposal, we will further optimize the peptides containing epitopes from OspC, FlaB and IR6 and will validate new immunoassays in both ELISA and rapid lateral flow point-of-care (POC) formats. An assay with greater specificity and improved sensitivity for detection of B. burgdorferi antibodies in the earliest stages of the disease, could lead to the development of a single-tier test capable of replacing the currently recommended two-tier paradigm. More importantly, given that early diagnosis and treatment of LD prevents illness progression and sequellae, the development of this test will make an important contribution to improvement of human health. PUBLIC HEALTH RELEVANCE: 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 sero- diagnostics 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-antigen peptide test. Because of greater specificity and improvement to the assay's sensitivity, this approach could lead to the development of a single tier assay to detect antibodies against any of the three pathogenic genospecies ofB. burgdorferi in both early and late Lyme disease.