间接免疫荧光法、免疫印迹法和多重微珠免疫法检测抗核抗体的比较和评价

Comparison and evaluate of indirect immunofluorescence,western blotting and multiple microbead immunoassay for detection of antinuclear antibodies

目的通过比较间接免疫荧光法、免疫印迹法和多重微珠免疫法三种方法检测抗核抗体的一致性,同时对免疫印迹法和多重微珠免疫法在检测抗核抗体谱的比较,评估各种方法的优势及其局限,为临床实践提供依据。方法收集578例同时进行三种方法检测的疑似自身免疫性疾病患者的血清样本检测结果,使用Kappa系数分析比较几种检测方法的一致性。结果在578例样本中,使用间接免疫荧光法检测抗核抗体阳性者280例(48.4%);使用免疫印迹法阳性者225例(38.9%);使用多重微珠免疫法阳性者252例(43.6%)。三种方法中任意两种方法检测抗核抗体的一致性均在85%以上,Kappa值均高于0.7,其中,间接免疫荧光法和免疫印迹法一致性最强,Kappa值为0.808(95%CI:0.761~0.855)。使用免疫印迹法与多重微珠免疫法检测9种抗核抗体谱,一致性均大于90%,而Kappa值则在0.535至0.839之间,其中一致性最强的抗体类型为抗SSA抗体和抗JO1抗体,分别为0.839和0.832。结论以上三种方法检测抗核抗体的一致性较高,其中间接免疫荧光法的阳性率最高,间接免疫荧光法和免疫印迹法一致性最强。免疫印迹法与多重微珠免疫法检测抗核抗体谱一致性较高。应联合使用三种方法,提高自身免疫性疾病的诊断率,避免漏检。

Objective By comparing the consistency of indirect immunofluorescence assay(IIFA),Immunoblot(IB),multiple microbead immunoassay(MBI) in the detection of antinuclear antibodies(ANA),and by comparing IB and MBI in the detection of ANA spectrum,the advantages and limitations of each method were evaluated to provide suggestions for clinical practice.Methods We collected serum samples from 578 patients who detected ANA by three methods since suspected of autoimmune disease.The consistency analysis of several methods was performed using the Kappa test.Results Among 578 serum samples,the numbers of ANA-positive samples were 280(48.4%),225(38.9%),and 252(43.6%) by IIFA,IB,and MBI,respectively.The consistency of any two methods was above 85%,and the Kappa values above0.7.Among them,the consistency of IIFA and IB was the strongest,and the Kappa value was 0.808(95% CI:0.761~0.855).For ANA spectrum detection,the consistency was higher than 90% by IB and MBI,and the Kappa values ranged form 0.535 and 0.839.The most consistent antibody types were anti-SSA antibody and anti-Jo1 antibody,which were 0.839 and 0.832,respectively.Conclusions The above three methods have high consistency in detecting antinuclear antibody,among which indirect immunofluorescence method has the highest positive rate,indirect immunofluorescence method has the strongest consistency with western blotting method,and western blotting method has high consistency with multiple microbead immunoassay for detecting antinuclear antibody.To improve the diagnosis of autoimmune diseases and avoid missed diagnosis,the three methods should be combined for ANA detection in clinical practice.