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[用免疫印迹法诊断莱姆病螺旋体病]

[Diagnosis of Lyme borreliosis with western blotting].

作者信息

Hulínská D

机构信息

Národní referencní laborator pro lymeskou boreliózu, Státní zdravotní ústav, Praha.

出版信息

Epidemiol Mikrobiol Imunol. 1997 Mar;46(1):3-8.

PMID:9162453
Abstract

Spirochetes Borrelia burgdorferi sensu lato, selected as antigens for Western blot analyses were isolated from cerebrospinal (strain 192 M) and from blood (strain Kc90) and identified by means of monoclonal antibodies and the polymerase chain reaction (PCR) as B. garinii and B. afzelii. Differences between B. garinii and B. afzelii are in the genotype of the surface protein OspA and OspB, internal flagellin (Fla II) and the main extracellular protein (MEP). The reaction of polyclonal antibodies in 918 serum specimens and 180 specimens of cerebrospinal fluid was investigated in IgG and IgM immunoblots in patients with neurological symptoms, arthritis and skin manifestations suspect of Lyme borreliosis. Confirmation of the immunoenzyme ELISA reaction by means of immunoblots in the acute stage of borreliosis, in clinically obscure cases, in herpetic infection, mononucleosis and leptospirosis revealed a higher sensitivity and specificity of Western blot. Proteins of B. garinii with a molecular weight of 94, 84, 66, 60, 56, 41, 39, 33, 29, 22, 18 and 14 kDa were detected in the reaction with monoclonal antibodies and immunoglobulins of patients suffering from barreliosis. The frequency and intensity of the reaction of these antigens differed markedly in sera of patients suffering from borreliosis and sera of patients who suffered from a different infection. The external surface antigen OspA, OspB, OspC and protein with 39 kDA are significant markers of borreliosis. The most frequently detected antigens in cross reactions with immunoglobulins against other pathogens are proteins P66, P60, P41 which are dominant immunogens of all types of borrelias and moreover a humoral response to them develops in the acute stage of the disease. In arthritis and neuroborreliosis a different in IgG immunoblots was found.

摘要

从脑脊液(192M株)和血液(Kc90株)中分离出作为蛋白质印迹分析抗原的疏螺旋体伯氏疏螺旋体狭义种,并通过单克隆抗体和聚合酶链反应(PCR)鉴定为加氏疏螺旋体和阿氏疏螺旋体。加氏疏螺旋体和阿氏疏螺旋体之间的差异在于表面蛋白OspA和OspB、内鞭毛蛋白(Fla II)和主要细胞外蛋白(MEP)的基因型。在疑似莱姆病疏螺旋体病的有神经症状、关节炎和皮肤表现的患者中,在IgG和IgM免疫印迹中研究了918份血清标本和180份脑脊液标本中多克隆抗体的反应。在疏螺旋体病急性期、临床隐匿病例、疱疹感染、单核细胞增多症和钩端螺旋体病中,通过免疫印迹法对免疫酶ELISA反应进行确认,结果显示蛋白质印迹法具有更高的敏感性和特异性。在与疏螺旋体病患者的单克隆抗体和免疫球蛋白反应中,检测到分子量为94、84、66、60、56、41、39、33、29、22、18和14 kDa的加氏疏螺旋体蛋白。这些抗原反应的频率和强度在疏螺旋体病患者血清和患有其他感染的患者血清中明显不同。外表面抗原OspA、OspB、OspC和39 kDa蛋白是疏螺旋体病的重要标志物。与针对其他病原体的免疫球蛋白发生交叉反应时最常检测到的抗原是P66、P60、P41蛋白,它们是所有类型疏螺旋体的主要免疫原,而且在疾病急性期会产生针对它们的体液反应。在关节炎和神经疏螺旋体病中,发现IgG免疫印迹存在差异。

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