间接酶联免疫法测定抗Jo-1抗体及其临床应用

Detection and clinical application of anti-histidyl-tRNA synthetase autoanbody by indirect enzyme-linked immunosorbent assay

目的利用基因工程表达的组氨酰转移核糖核酸合成酶自身抗原Jo-1蛋白,建立ELISA法,初步用于检测多发性肌炎/皮肌炎(PM/DM)中的抗Jo-1抗体。方法用表达蛋白建立间接ELISA,摸索反应时间、抗原包被浓度、样本稀释浓度及抗抗体工作浓度等工作条件,进一步检测方法的重复性及稳定性。用建立的方法,检测40例健康体检者、30例系统性红斑狼疮(SLE)、30例类风湿关节炎(RA)、20例干燥综合征(SS)、90例多发性肌炎/皮肌炎(PM/DM)患者血清中抗Jo-1抗体。结果成功建立间接ELISA法,并确立了一系列反应条件。临床检测结果显示,PM/DM患者抗Jo-1抗体的阳性率为25.6%,而非PM/DM患者均为阴性。PM/DM组Jo-1抗体阳性率与疾病对照组及健康对照组比较,差异均有统计学意义(均P<0.01)。结论本研究成功应用基因工程表达的Jo-1蛋白建立了间接ELISA法,检测抗Jo-1抗体,与国外报道的结果基本相当,与常用免疫印迹法对比检测的结果一致。

Objective To establish the enzyme linked immunosorbent assay （ELISA） to detect histidyl-tRNA synthetase autoantigen correlative antibodies for diagnosis of polymyotisis/dermatomsyositis. Methods The conditions of the enzyme linked immunosorbent assay （ELISA） with the fusion protein were optimized. The Jo-1 autoantigen correlative antibodies in serum of 90 patients with PM,/DM, 30 patients with SLE, 30 patients with RA, 20 patients with SS and 40 normal controls were detected by the established assay. Results When analyzing the serum of patients with PM/DM by ELISA, the result indicated the positive rate of anti-Jo-1 antibody was 25.6%, but the antibody was negative in other controls. There were significant differences of the positive autoantibody between PM/DM and disease controls or normal controls. Conclusions The indirect ELISA for anti-histidyl-tRNA synthetase autoantibody was established successfully. The results of ELISA showed similar positive rates, compared to reports in the literatures. The results of the established ELISA were consistent with those of the western blotting.