间接免疫荧光法与蛋白印迹法检测抗核抗体的比较研究

Comparison of Indirect Immunofluorescence and Line Immunoassay for Autoantibody Detection

目的 采用回顾性调查方法比较间接免疫荧光法（indirect immunofluorescence,IIF）与蛋白印迹法（line immunoassay,LIA）对抗核抗体检测的诊断价值.方法 采用间接免疫荧光法（IIF）与蛋白印迹法（LIA）对2 522份血清标本进行抗核抗体检测,检测结果可分为自身免疫病组（autoimmune disease,AID）、非AID组和非明确诊断组.并分析结果的相互关系和临床意义.结果 2 522份标本中,两种方法同时检测阳性752例,阳性率29.8%,显著高于单独IIF检测的阳性率24.3%（χ2=19.406,P＜0.01）和单独LIA检测的阳性率23.0%（χ2=30.18,P＜0.01）.IIF（＋）LIA（-）组荧光滴度分析表明,在滴度1∶100～1∶1 000组,非AID的阳性率显著高于AID组;而当滴度≥1∶1 000 AID阳性率显著高于非AID组（χ2=4.331,P＜0.05）;核型分布不同.IIF（-）/LIA 非阴性组蛋白印迹法显色强度与疾病的关系,显色越深,AID的可能性越高,15种已知抗体都有可能出现,抗Ro-52抗体出现频率最高.结论 该次分析结果表明IIF-ANA筛查和LIA-ANAs特异性抗体检测不能相互代替,临床应对需要通过检测ANA来排除AID患者的标本同时进行IIF-ANA的筛查和ANAs特异性抗体的检测,以避免仅用一种方法检测导致的AID患者漏诊.

Objective To evaluate the diagnostic performance of indirect immunofluorescence （IIF） and line immunoassay （LIA） for autoantibody （autoAb） detection by retrospective investigation method. Methods 2 522 cases of consecutive clin~ ical samples for ANA testing were tested by IIF and LIA for the detection of specific ANA antibodies. All the samples were divided into autoimmune diseases （AID） group, suspected AID group and non-AID group. The relationship between different test results and their clinical significance were analyzed. Results Of the 2 522 cases of specimens,752 cases （29.8%） were IIF-ANA＋/LIA-ANAs＋, higher than that （24.3%） of IIF-ANA ＋/LIA-ANAs -- （x2 = 19. 406, P〈0.01） and that （23.0 % ） of the IIF-ANA--/LIA-ANAs ＋ （23.0 % ） （x2= 30.18, P〈 0.01 ）. IIF （ ＋ ） LIA （ - ） group of fluorescence degree analysis showed that,in the drop of 1 ： 100~1 ： 1 000 the positive rate of non-AID group was higher than that of the AID and when the drop degree ≥1 ： 1 000 the positive rate of AID group was higher than that of the non-AID. The difference was statistically significan （x2 = 4. 331, P〈0.05） and the distribution of idiogram was different. In IIF （--）/LIA non nega- tive groups LIA color strength was relationship with disease,the deeper the color, the higher the possibility of AID. There were 15 kinds of known antibodies,and Ro-52 antibodies appear highest frequency. Conclusion The screening of ANA by IIF and the detection of ANA specific antibodies by LIA could not replace each other in clinical practice. In order to avoid misdiagnosis of patients with autoimmune diseases,it is recommend to screen ANA by IIF and detect ANA specific antibod- ies simultaneously when the ANA test is needed to excluded the presence of autoimmune diseases.