ASSAY TO DETECT ACUTE ACTIVE LYME DISEASE

莱姆病（LD）病例由于伯氏疏螺旋体（Bb）正在增加。检测活动性感染的试验是迫切需要的。本提案的主要目的是开发一种基于Bb特异性免疫复合物（IC）的活动性感染检测方法，供公众立即使用。有两个具体的目的来证明这一原则，并确认l）。在感染早期，BbIC先于游离Bb抗体（Ab）先阳性。BbIC表明活动性感染样本已经从LD患者中储存，这些患者具有病理学游走性红斑（EM）皮疹和通过Bb培养确认的微生物。基本原理是，在许多感染中，可以发现IC Ab比游离Ab更早地与其抗原（Ag）靶结合，并且与游离Ab相反，特异性IC Ab反映活动性感染。我们将在关键时间点研究系列设盲血清标本。将提供ELISA、免疫印迹和临床病史。BbIC将通过经证实的简单技术聚乙二醇分离。将通过免疫印迹和ELISA检查BbIC Ag和Ab。重点将放在特异性和体内表达的Bb Ag上。将对结果进行统计分析。第2阶段设计用于广泛使用和改进试验。拟议的商业应用：莱姆病活动性感染的准确实验室诊断是当务之急。每年进行超过500万次莱姆病血清学检测。无一例显示活动性感染。它们只显示过去的感染。一个测试，如伯氏疏螺旋体免疫复合物测定具有很高的潜力，作为一个标志物的活动性感染。它还可以比游离抗体检测更早地检测感染。因此，该检测方法可以满足莱姆病诊断的巨大需求，具有很高的市场潜力。

Lyme disease (LD) cases due to Borrelia burgdorferi (Bb) are increasing. Assays to detect active infection are urgently needed. The primary objective of this proposal is to develop an assay for active infection based upon Bb specific immune complexes (IC) for immediate public use. There are 2 Specific Aims to prove the principle and confirm that l). BbIC become positive before free Bb antibody (Ab) in early infection 2). BbIC indicate active infection Samples are already banked from LD patients with both the pathognomic erythema migrans (EM) rash AND microbiological confirmation by Bb cultures. The rationale is that in many infections IC Ab can be found bound to its antigen (Ag) target earlier than the free Ab, and in contrast to free Ab, specific IC Ab reflects active infection. We will study serial blinded serum specimens at key time points. ELISA, immunoblot and clinical histories will be available. BbIC will be isolated by a proven simple technique, polyethylene glycol. BbIC Ag and Ab will be examined by immunoblots and ELISA. Focus will be on specific and in vivo expressed Bb Ags. Results will be statistically analyzed. Phase 2 is designed for broadscale usage and refinement of the assay. PROPOSED COMMERCIAL APPLICATIONS: The need for accurate laboratory diagnosis of active infection in Lyme disease is imperative. There are over 5 million serologic Lyme disease tests performed per year. None indicate active infection. They only indicate past infection. A test such as the Borrelia burgdorferi immune complex assay has high potential as a marker of active infection. It can also detect infection early than free antibody assays. Therefore this assay can fulfill a great need in Lyme disease diagnosis and has high market potential.