摘要
目的 提取、半纯化RA33/ 36抗原 ,探讨RA33/ 36抗体对类风湿关节炎 (RA)诊断的特异性及敏感性。方法 以Ehrlich腹水癌细胞核提取物作粗抗原 ,采用肝素 琼脂糖凝胶CL - 6B层析柱对此粗抗原进行分离半纯化。分别用粗抗原和半纯化抗原作抗原 ,免疫印迹法检测 12 0例RA患者、40例红斑狼疮 (SLE)患者、35例干燥综合征 (SS)患者、2 5例系统性硬化症 (SSc)患者、10例混合性结缔组织病 (MCTD)患者和 30例正常对照血清中的RA33/ 36抗体。结果 ①SDS -PAGE示 2种抗原在相对分子质量 (Mr)为 33× 10 3 和 36× 10 3 处均可见条带 ,半纯化抗原杂带较粗抗原明显减少。②与粗抗原相比 ,半纯化抗原免疫印迹反应条带明显减少 ,且特别清晰 ,但阴性和阳性结果相同。③RA36抗体并不总是与RA33抗体以 2条带的形式同时出现。④RA33抗体和RA36抗体对RA诊断的敏感性差异无显著性 ,而特异性RA36抗体优于RA33抗体。⑤RA36抗体与RF、APF、ESR、关节相X线分期、关节功能分级无明显相关性。结论 半纯化抗原可直接用于检测RA33/ 36抗体 ,或用其作质量控制 ;RA36抗体可能是RA的一种可作为标记的特异性抗体 ,应对此抗体进行更深入研究。
Objective To extract and partial purify the RA33/36 antigen and to verify the diagnostic value of RA33/36 antibody for RA. Methods The RA33/36 antigen were partially purified from Ehrlich ascites carcinoma cell nuclear extracts by affinity chromatography on heparin sepharose CL 6B. Using the crude antigen and the partial purified antigen as antigenic sources on immunoblotting, we detected a total number of 260 sera from 120 patients with RA, 40 patients with SLE, 35 patients with SS, 25 patients with SSc, 10 patients with MCTD and 30 healthy controls. Results 1. The crude antigen and the partial purified antigen were observed as two bands of molecular weight of 33 000 and 36 000 in SDS PAGE according to the molecular weight marker. But the bands of the latter were obviously less than those of the former, and especially clear. 2. Compared with the crude antigen, the blot bands of the partial purified antigen were much significantly decreased and especially clear. But the positive rates had no difference between them. 3. RA36 antibody did not always appear with RA33 antibody. 4. There were no difference between the sensitivities of the two antibodies, but the specificity of RA36 antibody was higher than that of RA33 antibody. 5. The presence of anti RA36 was not correlated with RF, APF, ESR and clinical manifestations. Conclusion We should directly use the partial purified antigen to detect the antibodies in the future. The RA36 antibody may appear to be highly specific for RA, so we should make further studies on this antibody.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2001年第1期75-79,共5页
Chinese Journal of Microbiology and Immunology
基金
国家自然科学基金资助项目! (39770 70 3)
