酶联免疫吸附试验检测血清抗中性粒细胞胞质抗体方法的建立与临床应用

目的建立总抗原酶联免疫吸附试验(ELISA)检测血清抗中性粒细胞胞质抗体(ANCA)的方法,初步探讨其在ANCA相关血管炎中的表达及临床应用价值。方法从人的中性粒细胞中提取总抗原,用棋盘滴定法选择ELISA测定的最佳条件,确定临界值。测定系统性红斑狼疮(SLE)、肾病、血管炎、类风湿关节炎(RA)等患者血清总ANCA,并与间接免疫荧光(IIF)法、特异性髓过氧化物酶(MPO)-ANCA及蛋白酶3(PR3)-ANCA进行分析比较。结果总抗原ELISA与IIF法检测肾病、SLE、血管炎及RA患者血清ANCA阳性率比较差异均无统计学意义(P>0.05)。抗原特异性ELISA、IIF法检测肾病、血管炎、RA患者血清ANCA阳性率比较差异均无统计学意义(P>0.05),而SLE差异有统计学意义(P<0.05)。结论ELISA检测ANCA可行且具有广泛的推广应用前景。

Establishing a method to assay antineutrophil cytoplasmic antibody and its clinical application

Objective To establish a method of assaying antineutrophil cytoplasmic antibody(ANCA) in serum of patients with ANCA-associated vasculitis by total antigen enzyme-linked immunosorbent assay(ELISA). Methods Extract the total antigen from people′s neutrophil and choose the optimum condition of ELISA with chessboard typed titration and confirm the critical value .Assay the total ANCA in serum of patients with kidney disease,vasculitis,systemic lupus erythematosus(SLE) and rheumatoid arthritis(RA),compare with the methods of indirect immunofluorescence(IIF),myeloperoxidase(MPO)-ANCA and proteinase-3(PR3)-ANCA. Results The total ANCA in serum of four kinds of patients was assayed by total antigen ELISA and IIF method;the differences of ANCA positivity among the four kinds of patients showed no significance(P>0.05).The difference between antigen specificity ELISA and IIF had no statistical significance among the patients with kidney disease,vasculitis and RA,but SLE showed a statistical significance (P<0.05). Conclusion To assay ANCA by ELISA has extensive popularization and application prospects.