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首页> 外文期刊>Journal of Clinical Microbiology >Measurement of Antibody to Ureaplasma urealyticum by an Enzyme-Linked Immunosorbent Assay and Detection of Antibody Responses in Patients with Nongonococcal Urethritis
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Measurement of Antibody to Ureaplasma urealyticum by an Enzyme-Linked Immunosorbent Assay and Detection of Antibody Responses in Patients with Nongonococcal Urethritis

机译：酶联免疫吸附法测定解脲支原体抗体的含量，并检测非淋球菌性尿道炎患者的抗体应答

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The optimum conditions for the detection of human immunoglobulin G (IgG), IgM, and IgA antibodies to Ureaplasma urealyticum by an enzyme-linked immunosorbent assay (ELISA) were established by using a cell lysate antigen and commercially available alkaline phosphatase conjugates. No significant cross-reactions were observed among rabbit antisera to a variety of mycoplasmas of human origin and ureaplasma antigen, thus demonstrating the specificity of the ELISA. All human sera were assayed at a 1:200 dilution. Antigen was used at 20 μg of protein/ml and conjugates were diluted 1:500. Presence of IgG antibody to U. urealyticum was significantly associated with isolation of U. urealyticum (P < 0.001) in 110 women. Seventeen acute-phase and 19 convalescent-phase sera from male nongonococcal urethritis (NGU) patients were tested for the presence of antibody by both the metabolism inhibition assay and by ELISA, with overall agreements of 82 and 95% for acute- and convalescent-phase sera, respectively. Serum antibody responses were demonstrated to selected serotypes in the metabolism inhibition test, but the response as measured by the ELISA was independent of the serotype of the antigen used. Serum antibody levels in NGU patients were significantly higher (P < 0.002) than the normal serum standard in the IgG, IgM, and IgA classes. Additionally, the magnitude of change between acute- and convalescent-phase sera was greater for NGU patients than for normal asymptomatic ureaplasma-positive male controls. A significant change in antibody levels of one or more antibody classes was detected for 12 of 18 (67%) NGU patients by ELISA. Ten of the 12 (83%) individuals had a change in the IgM class, which is suggestive of an active infectious process. The ELISA is advantageous in that it requires only a single serotype antigen, uses one serum dilution, is class specific, and allows quantitative detection of differences between acute- and convalescent-phase sera.
机译：通过酶联免疫吸附测定法（ELISA）建立了用于检测解脲支原体的人免疫球蛋白G（IgG），IgM和IgA抗体的最佳条件，该酶可通过细胞裂解物抗原购得碱性磷酸酶结合物。在兔抗血清之间未观察到与多种人源支原体和脲原体抗原的显着交叉反应，从而证明了ELISA的特异性。所有人类血清均以1：200稀释度进行测定。抗原的使用量为20μg蛋白/ ml，缀合物按1：500稀释。存在针对 U的IgG抗体。解脲酶与 U的分离显着相关。 110名女性中的解脲支原体（ P <0.001）。通过代谢抑制试验和ELISA检测了男性非淋球菌性尿道炎（NGU）患者的17个急性期和19个恢复期血清的抗体存在情况，急性期和恢复期的总体一致性分别为82％和95％血清。在代谢抑制试验中，血清抗体应答被证明对选定的血清型，但是通过ELISA测定的应答与所用抗原的血清型无关。 NGU患者的血清抗体水平明显高于IgG，IgM和IgA类中的正常血清标准（ P <0.002）。此外，NGU患者的急性期和恢复期血清之间的变化幅度要比正常无症状脲原体阳性男性对照大。通过ELISA对18名（67％）NGU患者中的12名检测到一种或多种抗体类别的抗体水平有显着变化。 12名患者中有10名（83％）的IgM类别发生了变化，这表明感染过程活跃。 ELISA的优势在于，它仅需要一种血清型抗原，使用一种血清稀释液，具有特定的类别，并且可以定量检测急性期和恢复期血清之间的差异。

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