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以尼龙为固相的酶联免疫吸附试验检测系统性红斑狼疮中可提取核抗原的抗体

Enzyme-linked immunosorbent assay for detection of antibodies to extractable nuclear antigens in systemic lupus erythematosus, with nylon as solid phase.

作者信息

Ishaq M, Ali R

出版信息

Clin Chem. 1983 May;29(5):823-7.

PMID:6839459
Abstract

In this enzyme-linked immunosorbent assay (ELISA) for detection of antibodies against extractable nuclear antigens (ENA) in sera of patients with systemic lupus erythematosus (SLE), nylon is used as solid phase for antigen binding instead of the commonly used polystyrene surface. Optimal conditions for activation of the nylon beads, antigen coating, and other relevant factors have been investigated. We compared the incidence of anti-ENA antibodies in SLE, using chromogenic and fluorogenic enzyme substrates. Of SLE patients, 54% were positive for anti-ENA antibodies when chromogenic substrate was used as compared with 68% for fluorogenic substrate. Antibody activity against Sm and RNP antigens was distinguished on the basis of ribonuclease sensitivity of the RNP antigen. The method described offers advantages such as decreased background activity, increased surface area, facility for prolonged storage of antigen-coated solid phase, and miniaturization of the assay.

摘要

在这种用于检测系统性红斑狼疮(SLE)患者血清中抗可提取核抗原(ENA)抗体的酶联免疫吸附测定(ELISA)中,尼龙被用作抗原结合的固相,而非常用的聚苯乙烯表面。已对尼龙珠激活、抗原包被及其他相关因素的最佳条件进行了研究。我们使用显色和荧光酶底物比较了SLE患者中抗ENA抗体的发生率。当使用显色底物时,54%的SLE患者抗ENA抗体呈阳性,而使用荧光底物时这一比例为68%。基于RNP抗原的核糖核酸酶敏感性区分了针对Sm和RNP抗原的抗体活性。所描述的方法具有以下优点:背景活性降低、表面积增加、抗原包被固相便于长期保存以及检测小型化。

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