以鸡卵核42000蛋白为抗原的斑点金免疫渗滤法的建立及对SLE血清学诊断的评价  

Development and evaluation of a dot immunogold filtration assay for the serodiagnosis of systemic lupus erythematosus using 42-kD nuclear protein of chicken egg as antigen

作　　者：颜丹[1] 李燎[1] 陈德宇[1] 钟桂书[1] 胡筠惠[2] 钟建泉[3] 

机构地区：[1]四川泸州医学院附属医院皮肤科,646000 [2]泸州医学院附属卫生学校 [3]四川泸州医学院附属医院检验科,646000

出　　处：《中华皮肤科杂志》2007年第1期28-30,共3页Chinese Journal of Dermatology

摘　　要：目的开发一种基于斑点金免疫渗滤法的系统性红斑狼疮（SEE）血清学诊断方法。方法我们先前的研究提示抗鸡卵核42000蛋白可与大部分SLE患者中的自身抗体特异性反应，本研究以该蛋白为待测抗原建立斑点金免疫渗滤法（DIGFA），并检测了191例治疗前后的SLE患者血清、198例结缔组织病患者血清（皮肌炎，多发性肌炎72例，硬皮病48例。混合性结缔组织病78例）和100例健康对照者血清。以抗鸡卵核42000蛋白特异性IgM和IgG的几何平均滴度倒数（GMRT）为指标评价其对SLE血清学诊断的有效性。结果本方法用于SLE血清学诊断的敏感性和特异性分别为94．24％（180／191 SLE患者血清呈阳性反应）和100．00％（所检测的皮肌炎／多发性肌炎、硬皮病、混合结缔组织病及健康对照者血清均呈阴性反应）。在174例（71例活动期，103例非活动期）特异性IgM和IgG抗体均阳性的SLE患者中，活动期与非活动期患者治疗前后IgM、IgG的GMRT均显著下降（P值均〈0．01）。且IgM的GMRT下降幅度较IgG的更大。结论以鸡卵核42000蛋白为抗原的斑点金免疫渗滤法对SLE血清学诊断敏感特异，抗鸡卵核42000蛋白特异性IgM水平的检测在疗效评定方面优于对相应IgG的检测。Objective To develop a new serodiagnostic method for systemic lupus erythematosus （SLE） using a dot immunogold filtration assay （ DIGFA ）. Methods The 42-kD egg nuclear protein, which was proved to specifically react with autoantibodies in the sera of many patients with SLE in our previous study, was separated from the chicken eggs by SDS-PAGE and analyzed by Dot-ELISA. A dot immunogold filtration assay using 42-kD nuclear protein of chicken egg as antigen was developed. Sera from patients with SLE before and after treatment （ n = 191 ）, dermatomyositis or polymyositis （ DM/PM ） （ n = 72 ）, progressive systemic sclerosis （ PSS ） （ n = 48 ）, mixed connective disease （ MCTD ） （ n = 78 ）, and healthy control subjects （ n = 100 ） were assayed by the established DIGFA. The primary evaluation parameter was the geometric mean reciprocal titer （ GMRT ） of specific IgM and IgG. Results The sensitivity and specificity of the established DIGFA were 94.24% （ 180 of 191 SLE sera showed a positive result ） and 100.00% （ all sera of healthy controls, patients with DM/PM, PSS, and MCTD showed a negative result ）, respectively. Of the 174 SLE patients with both specific anti±2-kD nuclear protein IgM and IgG antibody in the sera, 71 were with active SLE and 103 were with inactive SLE. In those with active SLE, the specific IgM and IgG in sera before treatment showed significant higher GMRT than after treatment （ 768.84 ± 26.43 vs 367.53 ± 18.52, 629.25 ± 30.05 vs 405.72 ± 16.35, respectively.） （ P 〈 0.01 ）. Meanwhile, in those with inactive SLE, the GMRT of specific IgM and IgG was also significantly reduced after treatment （421.34 ± 23.12 to 295.63 ± 20.57, 452.67 ± 16.35 to 358.73 ± 21.27, respectively） （ P〈 0.01 ）. Conclusions The results suggest that DIGFA using 42-kD nuclear protein of chicken egg as antigen is a valuable tool for the serodiagnosis of systemic lupus erythematosus, and GMRT of anti-42 kD nuclear protein-specific IgM determined by

关 键 词：红斑狼疮 系统性 斑点金免疫渗滤法 鸡卵核42000蛋白 血清学诊断 

分 类 号：R686[医药卫生—骨科学]