IMMUNODOTS & ELISA WITH RECOMBINANT BORRELIA BURGDORFERI

莱姆病的早期诊断和治疗与 改善临床结局，并建议预防后续 表现为关节炎和颅麻痹。 不幸的是， 莱姆病的早期诊断受到阻碍，因为B.burgdorferi 抗原和人类对它们的体液应答仅部分地被 特征，并且因为没有可靠的血清学测定， 疾病的诊断。 我们建议发展免疫斑点和ELISA检测来诊断莱姆病 基于克隆B的疾病。人类的伯氏菌基因产物 免疫原 我们将研究这些的敏感性和特异性。 诊断分析 B。将使用的伯氏菌基因产物是 不同于所有表征的抗原如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 从在E.杆菌 只与莱姆病患者的血清反应， 与抗OspA/OspB、抗OspC或抗鞭毛蛋白抗体反应，或 与正常对照的血清。 我们会完成DNA测序 这个克隆，表征其基因产物的生物化学， 净化他们。 重组B。将分析伯氏抗原， ELISA和免疫斑点结合试验用于区分 来自莱姆病患者的血清，来自莱姆病患者的血清， 其他传染病和正常人的血清。

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.