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[利用纯化抗原开发抗梅毒螺旋体-IgM抗体检测方法——与传统方法的比较]

[Development of anti-Treponema pallidum-IgM antibodies detection methods using purified antigen--comparison with conventional methods].

作者信息

Sato T

机构信息

Department of Microbiology, Yamaguchi University School of Medicine.

出版信息

Kansenshogaku Zasshi. 1993 Aug;67(8):758-66. doi: 10.11150/kansenshogakuzasshi1970.67.758.

Abstract

Treponema pallidum subsp. pallidum Nichols (Tp) antigens were purified by centrifugation in the presence of Hypaque (sodium diatrizoate) or MgCl2. When the TPHA (Treponema pallidum hemagglutination) tests with crude and purified antigens were carried out in the sera of patients with untreated primary syphilis, the purification by MgCl2 resulted in enhanced sensitivity to Tp-specific IgM as the same as the purification by Hypaque. In addition, the SPHA (solid phase hemadsorption) test with the Hypaque-purified antigen to anti-Tp-IgM antibodies was carried out using the sera of patients with primary and secondary syphilis. Fifty-nine percent (27/46) of them showed positive for the crude antigen and 100% (46/46) for the purified antigen, indicating that sensitivity to anti Tp-IgM was clearly enhanced with the antigen purification. The captured Tp-IgM-RIA test with the purified antigen was also undertaken in 16 patients with primary syphilis, 31 patients with secondary syphilis and 73 patients with latent syphilis. All the patients in the former two groups were positive, while all of the patients in the latter group were negative. To elucidate the mechanism whereby the purification improves the sensitivity to anti-Tp-IgM antibodies, host-derived antibodies associated with crude and purified Tp cells were measured by a fluorescent antibody technique. As the results, a great amount of rabbit anti-Tp-IgM antibodies were found on the surface of the crude antigen, whereas only a trace amount was detected on the purified antigens.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

梅毒螺旋体苍白亚种Nichols株(Tp)抗原通过在泛影葡胺(泛影酸钠)或MgCl₂存在下离心进行纯化。当用粗制抗原和纯化抗原对未经治疗的一期梅毒患者血清进行梅毒螺旋体血细胞凝集试验(TPHA)时,MgCl₂纯化与泛影葡胺纯化一样,提高了对Tp特异性IgM的敏感性。此外,使用一期和二期梅毒患者的血清,对经泛影葡胺纯化的抗原进行抗Tp-IgM抗体的固相血细胞吸附试验(SPHA)。其中59%(27/46)的患者粗制抗原检测呈阳性,100%(46/46)的患者纯化抗原检测呈阳性,表明抗原纯化明显提高了对抗Tp-IgM的敏感性。还对16例一期梅毒患者、31例二期梅毒患者和73例潜伏梅毒患者进行了用纯化抗原的捕获Tp-IgM-RIA试验。前两组的所有患者均为阳性,而最后一组的所有患者均为阴性。为阐明纯化提高对抗Tp-IgM抗体敏感性的机制,采用荧光抗体技术检测了与粗制和纯化Tp细胞相关的宿主来源抗体。结果发现,大量兔抗Tp-IgM抗体存在于粗制抗原表面,而纯化抗原上仅检测到微量。(摘要截于250字)

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