鸡卵核中72000蛋白的纯化及其对混合性结缔组织病的诊断价值

PURIFICATION OF PROTEIN-72000 FROM EGG NUCLEUS AND ITS APPLICATION TO DIAGNOSIS OF PATIENTS WITH MIXED CONNECTIVE TISSUE DISEASE

目的 :纯化鸡卵核中 72 0 0 0蛋白 ,并探讨该抗原对混合性结缔组织病 (MCTD)的诊断价值。方法 :以常规SDS -PAGE和电洗脱技术相结合 ,分离纯化鸡卵核中 72 0 0 0蛋白 ,并用于Dot-ELISA诊断MCTD。结果 :用该技术分离纯化了鸡卵核中 72 0 0 0蛋白 ,获得约 2 0 0 0 μg具有明显免疫反应性的高纯度抗原。用于检测MCTD病人血清78份 ,阳性 6 3份 ,阳性反应率为 80 .77% ;正常人血清 10 0份未发现假阳性 ;对皮肌炎和多发性肌炎 (DM PM) 72份 ,硬皮病 (PSS) 4 8份和系统性红斑狼疮 (SLE) 12 6份血清亦无交叉反应出现。结论 :以纯化的鸡卵核中 72 0 0 0蛋白为抗原 ,建立的Dot-ELISA方法 。

Objective: To purify the 72000 protein from egg nucleus,and to study its value in diagnosis of patients with mixed connective tissue disease (MCTD).Methods: The 72000 protein from egg nucleus was extracted and then purified by common SDS-PAGE and electroelution technique,and was applied for diagnosing MCTD with Dot-ELISA.Results: The 72000 protein was purified by these techniques, about 200μg of purified antigen was obtained which showed apparent immunogenicity and high purity.When it was applied for diagnosis,the positive rate was 80.77%(63/78)in case of MCTD,but no false positive and cross reaction were found in sera from 100 normal individuals,72 sera of dermatomyositis or polymyositis (DM/PM),48 sera of progressed systemic scleroderma (PSS) and 126 sera of systemic lupus erythematosus (SLE). Conclusion:The purified 72000 antigen from egg nucleus as a diagnostic antigen in MCTD by means of Dot-ELISA is considered favorably as specific and sensitive.