IMMUNODOTS & ELISA WITH RECOMBINANT BORRELIA BURGDORFERI

莱姆病的早期诊断和治疗与 改善临床结果，并建议防止后续 表现为关节炎和颅瘫。不幸的是， 莱姆病的早期诊断一直受到阻碍，因为伯氏杆菌 抗原和人类对它们的体液反应只是部分 特征，因为目前还没有可靠的血清学检测方法 疾病的诊断。 我们建议建立免疫斑点和酶联免疫吸附试验来诊断莱姆病。 基于克隆的人类伯氏杆菌基因产物的疾病 免疫原。我们将调查这些指标的敏感性和特异性。 诊断化验。拟使用的伯氏杆菌基因产物为 区别于所有特征抗原，如OspA/OSPB、OspC、鞭毛蛋白 和热休克蛋白。在初步研究中，我们发现了一种 伯氏杆菌lambdagt11B.burgdorferi文库在大肠杆菌中的表达 这只与莱姆病患者的血清发生反应，但失败了 与抗OspA/OSPB、抗OspC或抗鞭毛抗体或 取自正常对照的血清。我们将完成DNA的测序 这个克隆，对它的基因产物进行了生化表征，并部分地 净化它们。重组伯氏杆菌抗原将在年进行分析 酶联免疫吸附试验和免疫斑点结合试验检测其鉴别能力 莱姆病患者的血清与莱姆病患者的血清之间 其他传染病，以及正常人的血清。

Early diagnosis and treatment of Lyme disease is associated with an improved clinical outcome, and is recommended to prevent subsequent manifestations such as arthritis and cranial paralysis. Unfortunately, early diagnosis of Lyme disease has been hampered because B.burgdorferi antigens and the human humoral response to them have been only partially characterized, and because there are no reliable serological assays for diagnosis of the disease. We propose to develop immunodots and ELISA assays for diagnosis of Lyme disease based on cloned B. burgdorferi gene products that are human immunogens. We shall investigate sensitivity and specificity of these diagnostic assays. The B. burgdorferi gene-products to be used are distinct from all characterized antigens as OspA/OspB, OspC, flagellin and heat-shock proteins. In preliminary studies, we have identified a single clone from a lambdagt11 B.burgdorferi library expressed in E. coli that reacted only with sera from patients with Lyme disease and failed to react with anti-OspA/OspB, anti-OspC or anti-flagellin antibodies or with sera from normal controls. We will finish sequencing the DNA from this clone, characterize its gene-products biochemically, and partially purify them. Recombinant B. burgdorferi antigens will be analyzed in ELISA and immunodot binding assays for their ability to discriminate between sera from patients with Lyme disease, sera from patients with other infectious diseases, and sera from normal individuals.