Multi-antigen peptide assay for the serodiagnosis of Lyme disease

描述（由申请人提供）：莱姆病（LD）是美国最常见的病媒传染病，仍然是一个重大的公共卫生问题。LD的实验室诊断取决于对伯氏疏螺旋体抗体的证明。目前的血清学检测在早期LD中特异性不够，敏感性不高。需要新的检测方法，我们计划填补这一未满足的需求。我们的主要目标是生产新的敏感和特异性的肽为基础的检测LD的血清学诊断。在第一阶段，我们在定义新的血清学测定的组分方面取得了很好的进展：我们开发了IR6肽，其具有更广泛的检测表达不同VlsE序列的螺旋体感染的患者中的抗体反应性的能力;在17个已知的US OspC组中，我们已经鉴定了4个（B、F、I和K）作为表位作图的潜在靶标;并且我们评估了由来自OspC（10个残基）、FlaB（13个残基）和VlsE-IR6（17个残基）的序列组成的多抗原肽结合来自患有LD的个体的抗体的能力。在这个II期提案中，我们将进一步优化含有OspC、FlaB和IR6表位的肽，并将在ELISA和快速侧流床旁（POC）模式中验证新的免疫测定。一种检测B的特异性更强、灵敏度更高的检测方法。在疾病的最早阶段检测伯氏抗体，可能导致能够取代目前推荐的两级范式的单层测试的发展。更重要的是，鉴于LD的早期诊断和治疗可以防止疾病进展和后遗症，该测试的开发将为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病媒传染病，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抗原肽检测取代目前的检测方法。由于更高的特异性和提高检测的灵敏度，这种方法可能导致一个单一的层次检测的发展，以检测抗体对任何三个致病基因种的B。莱姆病的早期和晚期都有burgdorferi。

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.