流式荧光免疫技术检测抗核抗体谱的性能与临床应用评价  

The Performance and Clinical Application Evaluation of Flow CytometryFluorescence Immunoassay for Detecting AntinuclearAntibody Spectrum

作　　者：李晓宁 李丽娟[1] 侯艳峰 王泽霖 闫存玲[1] 李志艳[1] LI Xiaoning;LI Lijuan;HOU Yanfeng;WANG Zelin;YAN Cunling;LI Zhiyan(Department of Laboratory Medicine,Peking University First Hospital,Beijing 100034,China;St Francis Methodist School,Singapore)

机构地区：[1]北京大学第一医院检验科,北京100034 [2]圣法西斯卫理学校,新加坡

出　　处：《标记免疫分析与临床》2023年第11期1894-1898,1971,共6页Labeled Immunoassays and Clinical Medicine

摘　　要：目的分析流式荧光免疫技术(FFIA)检测自身抗体谱的结果及评价其在系统性红斑狼疮(SLE)诊断中的应用价值。方法用流式荧光免疫法和免疫印迹法同时检测219例自身免疫病患者(包括92例SLE患者和127例非SLE的其他自身免疫病患者)、231例其他疾病患者、50例健康体检者血清的特异性自身抗体(抗nRNP、Sm、SS-A、SS-B、Jo-1、Scl-70、rRNP、PCNA、Nucleosome、Histone、CenP-B、M2、dsDNA),计算两种方法的阴阳符合率,并采用Kappa检验评估结果一致性;采用受试者工作特征曲线(ROC)比较各抗体在系统性红斑狼疮诊断中的应用价值。结果FFIA与免疫印迹法检测各项自身抗体总符合率在87%至98%之间,Kappa值在0.179至0.854之间,其中抗CenP-B抗体一致性最好(Kappa值=0.854),抗His抗体一致性最差(Kappa值=0.179)。在219例自身免疫病例组数据中,根据试剂盒给定的阳性结果判断标准得到的免疫印记法灵敏度(62.0%)高于流式荧光(44.6%),而特异性略低于后者,分别为89.8%和94.4%,但二者诊断符合率基本一致且较好,分别为80.3%和77.3%。在用流式荧光免疫法检测各抗体用于SLE诊断的效能比较中,绘制ROC曲线结果显示抗dsDNA抗体对SLE诊断价值最高,AUC为0.768,其最佳cut-off值为10.94IU/mL,此时对SLE和非SLE自身免疫性疾病判断的灵敏度为65.2%,特异性87.4%,总符合率为78.1%,其他各项在最佳cut-off值时,抗Sm抗体的特异性(92.1%)最高,但灵敏度(28.3%)较差,抗CenP-B抗体的特异性(16.5%)最差,但灵敏度(100%)最高。结论FFIA检测抗核抗体谱与免疫印迹技术相比,一致性尚可,阴性符合率较阳性符合率更高,总符合率较好。FFIA检测抗dsDNA抗体、抗rRNP抗体、抗nRNP抗体、抗Nucl抗体和抗Sm抗体用于SLE诊断效能较好,且FFIA可实现自动化、大规模化、高效化检测。实验室在日常工作中可结合自身工作量、临床需求等选择适合本实验室流程的方法学。Objective To analyze the results of flow cytometry fluorescence immunoassay(FFIA)in detecting autoantibody profiles and to evaluate its application value in the diagnosis of systemic lupus erythematosus(SLE).Methods The specific autoantibodies(anti-nRNP,Sm,SS-A,SS-B,Jo-1,Scl-70,rRNP,PCNA,Nucleosome,Histone,Cenp-B,M2,dsDNA antibody)in serum samples of 219 patients with autoimmune diseases(including 92 SLE patients and 127 non SLE patients with other autoimmune diseases),231 patients with other diseases,and 50 healthy individuals were detected simultaneously using flow cytometry and immunoblotting methods.The coincidence rate of the two methods was calculated,and the consistency of the results was evaluated by Kappa test.Receiver operating characteristic curve(ROC)was drawn to compare the application value of antibodies in the diagnosis of systemic lupus erythematosus.Results The total coincidence rate between FFIA and immunoblotting for detecting various autoantibodies ranged from 87%to 98%,with Kappa values ranging from 0.179 to 0.854.Among them,the consistency of anti-CenP-B antibodies was the best(Kappa value=0.854),while the consistency of anti-His antibodies was the worst(Kappa value=0.179).In the data of 219 autoimmune case groups,the sensitivity(62.0%)of immunoblotting based on the positive result judgment criteria given by the reagent kit was higher than that of flow cytometry(44.6%),while the specificity was slightly lower than the latter,with values of 89.8%and 94.4%,respectively.However,the diagnostic accuracy of the two methods was generally the same and decent,with 80.3%and 77.3%,respectively.In the comparison of the efficacy of using flow cytometry fluorescence immunoassay to detect various antibodies for SLE diagnosis,ROC curve results showed that anti-dsDNA antibodies had the highest diagnostic value for SLE,with an AUC of 0.768 and an optimal cut-off value of 10.94 IU/mL.The sensitivity for judging SLE and non SLE autoimmune diseases was 65.2%,while the specificity was 87.4%,and the total coincide

关 键 词：抗核抗体谱 流式荧光免疫法 免疫印迹技法 系统性红斑狼疮 

分 类 号：R593.241[医药卫生—内科学]