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。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2个具体的目的来证明这一原则，并证实1)。在感染早期游离Bb抗体（Ab）前，BbIC呈阳性。BbIC表明活动性感染的样本已经从LD患者中保存下来，这些患者既有病征性移动性红斑（EM）皮疹，也有Bb培养的微生物学证实。其基本原理是，在许多感染中，可以发现IC Ab比游离Ab更早地与抗原（Ag）靶标结合，与游离Ab相比，特异性IC Ab反映了活动性感染。我们将在关键时间点研究连续盲法血清标本。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——聚乙二醇进行分离。通过免疫印迹和ELISA检测血清抗原和抗体。重点将放在特异性和体内表达的Bb Ags上。结果将进行统计分析。第2阶段旨在广泛使用和改进该检测方法。建议的商业应用：对莱姆病活动性感染的准确实验室诊断是迫切需要的。每年进行500多万例莱姆病血清学检测。无表示活动性感染。它们只表明过去感染过。像伯氏疏螺旋体免疫复合体试验这样的测试作为活动性感染的标记物具有很高的潜力。它还可以比自由抗体测定法更早地发现感染。因此，该方法可以满足莱姆病诊断的需要，具有很大的市场潜力。

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.