ASSAY TO DETECT ACUTE ACTIVE LYME DISEASE

由伯氏疏螺旋体(BB)引起的莱姆病(LD)病例正在增加。迫切需要检测活动性感染的检测方法。这项建议的主要目标是开发一种基于BB特异性免疫复合体(IC)的活动性感染检测方法，立即供公众使用。有两个具体的目的来证明原理和确认L)。在感染早期，BbIC在游离BB抗体(Ab)前转为阳性。BbIC表明，活动性感染样本已经来自患有病理性移行性红斑(EM)皮疹且经BB培养确认的LD患者。其基本原理是，在许多感染中，可以发现IC抗体比游离抗体更早与其抗原(Ag)靶标结合，而与游离抗体相比，特异性IC抗体反映了活动性感染。我们将研究关键时间点的连续盲法血清样本。将提供ELISA、免疫印迹和临床病史。BbIC将通过一种经过验证的简单技术-聚乙二醇法进行分离。用免疫印迹法和酶联免疫吸附试验检测BbIC抗原和抗体。重点将放在特定的和体内表达的BB AGS上。结果将进行统计分析。第二阶段是为广泛使用和分析的改进而设计的。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。每年进行的莱姆病血清学检测超过500万次。没有一例表明有活动性感染。它们只表明过去感染过。伯氏疏螺旋体免疫复合体检测作为活动性感染的标志物具有很高的潜力。它还可以比游离抗体检测更早地发现感染。因此，该方法能很好地满足莱姆病诊断的需要，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.