应用蛋白质芯片技术检测血清自身抗体的初步探讨

目的研究应用蛋白质与抗原分子微阵列技术检测血清自身抗体。方法运用胶体金免疫金银染色法在氧化型琼脂糖凝胶膜结合戊二醛(GA)分子衍生的活性表面,制作蛋白质与抗原分子微阵列,建立检测血清自身抗体的方法。结果在未经选择的自身免疫病患者中[包括系统性红斑狼疮(SLE)、类风湿关节炎(RA)和混合性结缔组织病(MCTD)],共检出有多种自身抗体阳性,其中抗ssDNA抗体、抗甲状腺球蛋白(TG)抗体、抗甲状腺过氧化切酶(TPO)抗体、子宫内膜抗体(EmAb)和类风湿因子(RF)等的检出频率较高;但心磷脂抗体(ACA)、抗肝细胞膜特异性脂蛋白(LSP)和抗精子抗体(AsAb)未检出1项阳性,经初步的方法学对比考证,与酶联免疫吸附试验(ELISA)测试结果有较好的重复性(结果一致性达90%)。结论本法敏感度高,结果直观且实验条件易于控制,为阵列芯片的可视化提供了一种很好的模式。

Using protein and antigen microarray technique for autoantibody analysis

Objective Using the protein and antigen microarray technique based on immune gold silver staining to analyze autoantibodies. Methods Based on molecular assembly technique,the matrix of agarose gel was deposited on the surface of glass plate,followed by drying,oxidation with NaIO4 and cross linking with glutaraldehyde.The protien and antigen microarry was fabricated,and immune gold silver staining(IGSS) immunoassay for autoantibodies was developed on this microarray. Results Many kinds of autoantibodies were positive in patients with autoimmune diseases,including systemic lupus erythematosus(SLE),reumatoid arthritis(RA) and mixed connective tissue disease(MCTD).Among them,there was higher positive rate of autoantibodies to ssDNA,thyroglobulin,thyroid peroxidase,endometrial and AgRIgG,but negative of autoantibodies to cardiolipin,LSP and spermatozoa.In comparison with the results of enzyme-linked immunosorbent assay(ELISA),this technique had a good reproducibility(90% of results of both methods were identical.). Conclusion This technique is sensitive,eyeable and easy controlled,and provides a better way of experiment for further investigating an visible array-chip.