抗瓜氨酸化蛋白抗体特异性免疫复合物ELISA法的建立与临床初步应用  

Development and clinical preliminary application of ELISA for anti-citrullinated protein antibody specific immune complexes

作　　者：李晓军[1] 王凯[1] 高德玉[1] 虞伟[1] 严孝岭[1] 陈芳芳[1] 刘阳[1] 

机构地区：[1]南京军区南京总医院临床中心实验科,南京210002

出　　处：《临床检验杂志》2014年第3期177-180,共4页Chinese Journal of Clinical Laboratory Science

基　　金：国家自然科学基金项目(81071419);江苏省社会发展基金(BE2011786)

摘　　要：目的建立检测血清抗瓜氨酸化蛋白抗体特异性免疫复合物(ACPA-IC)的ELISA技术,探讨ACPA-IC对类风湿关节炎(RA)的临床意义。方法建立检测血清ACPA-IC的ELISA技术,并对方法的特异性、稳定性和重复性进行评估。采用PEG6000沉淀法自RA患者血清中提取IC。用建立的ACPA-IC ELISA法对92例RA患者包括46例ACPA阳性(ACPA+)患者和46例ACPA阴性(ACPA-)患者、46例SLE患者及47例健康献血员血清ACPA-IC水平进行检测。并分析RA患者血清ACPA-IC水平与ACPA和类风湿因子(RF)的相关性。结果经方法学评估,建立的ACPA-IC ELISA法在特异性、稳定性和可重复性方面均达到检测要求。ACPA+患者ACPA-IC水平高于ACPA-患者、SLE患者及健康对照组(P<0.01)。当cut off值设为0.9时,该法测得RA患者血清ACPA-IC的阳性率为45.65%(42/92),显著高于SLE(6/46,13.04%)及健康对照组(2/47,4.26%),χ2分别为14.376和24.632,P均<0.01。46例ACPA+RA患者ACPA-IC阳性者36例(78.26%),而ACPARA患者ACPA-IC阳性者仅6例(13.04%),差异有统计学意义(χ2=39.429,P<0.01)。ACPA-IC与ACPA和RF的Spearman相关分析结果表明,RA患者血清ACPA-IC水平分别与血清ACPA(r=0.708,P=0.000)、RF(r=0.571,P=0.000)呈正相关。结论成功建立检测血清ACPA-IC水平的ELISA技术。RA患者尤其是ACPA+患者血清ACPA-IC水平显著升高。ACPA-IC在RA患者血清中出现可能具有重要的致病意义。Objective To establish an ELISA for detecting serum anti-citrullinated protein antibody specific immune complex （ACPA- IC） in serum, and investigate its clinical signlficances in rheumatoid arthritis （RA） patients. Methods The ELISA was developed for measuring the serum level of ACPA-IC, and the specificity, stability and reproducibility of the assay were evaluated. The immune com- plex in serum was isolated with PEG6000 precipitation. Ninety-two patients with RA, including 46 ACPA positive RA patients and 46 ACPA negative RA patients, 46 SLE patients and 47 healthy controls were investigated. The levels of serum ACPA-IC of the three groups were determined by the established method and the correlations between ACPA-IC and ACPA, rheumatoid factor （RF） were an- alyzed respectively. Results The specificity, stability, and reproducibility of the assay fulfilled the requests of clinical testing by methodological evaluation. The levels of serum ACPA-IC in ACPA positive RA patients were significant higher than those in ACPA neg- ative RA patients, SLE patients and healthy controls （P 〈0. O1 ）. When cut off value for the ELISA was set at 0.9, the positive rate of ACPA-IC in RA patients （45.65%, 42/92） was significantly higher than those in SLE （6/46,13.04%） and healthy controls （2/47, 4.26% ） （X2 = 14.376, 24.632,P 〈0.01 ）. In the 46 ACPA positive RA patients, 36（78.26% ） were ACPA-IC positive, but only 6（ 13.04% ） were positive in 46 ACPA negative RA patients. There was significant difference of ACPA-IC positive rate be- tween the ACPA-positive and negative RA patients（x2 = 39.429, P 〈 0.01 ）. The positive correlation existed between ACPA-IC and ACPA（r =0.708, P 〈0.001） or RF（r=0.571, P〈0.001） in the RA patients. Conclusion We developed a novel ELISA for de- tecting serum ACPA-IC. Serum ACPA-IC levels in RA patients especially ACPA positive RA patients were significantly elevated. ACPA-IC in serum of RA patients may play important pathogenic significance.

关 键 词：抗瓜氨酸化蛋白抗体 免疫复合物 类风湿关节炎 酶联免疫吸附试验 类风湿因子 

分 类 号：R446.5[医药卫生—诊断学]