目的：采用酶联免疫吸附试验（enzyme-linked immunosorbent assay, ELISA）, 观察抗双链DNA（double-stranded DNA, dsDNA）IgG抗体在系统性红斑狼疮（systemic lupus erythematosus, SLE）患者和非SLE患者中的分布特点, 并评估ELISA法联合绿蝇短膜虫间接免疫荧光试验（crithidia luciliae immunofluorescence test, CLIFT）检测抗dsDNA-IgG抗体用于SLE诊断的效能。方法：收集上海交通大学医学院附属瑞金医院检测抗dsDNA-IgG抗体的住院患者的临床资料, 共4 726例采用ELISA法, 其中830例同时采用CLIFT检测, 用受试者工作特征曲线（receiver operating characteristic curve, ROC）分析2种检测方法辅助诊断SLE的效能。结果：采用ELISA法检测的 4 726例住院患者中, dsDNA抗体阳性者398例, 其中SLE 183例, 占dsDNA 阳性者46.0%, 非 SLE疾病依次为肝脏疾病（28.4%）、其他结缔组织病（7.5%）、血液系统疾病（5.3%）及呼吸系统疾病（3.8%）;830例采用CLIFT检测ds DNA, 134例结果显示为dsDNA抗体阳性的患者中, 其中SLE者占94.0%, 另有非SLE诊断病例包括自身免疫性肝病3例, 肝硬化、肾炎、银屑病和抗磷脂综合征各1例。830例完成ELISA法与CLIFT双检测的患者中, SLE患者共197例, 非SLE患者633例, ELISA法检测灵敏度为74.6%, 特异度仅为92.9%;而CLIFT法测dsDNA抗体具有较高的特异度, 为98.7%, 灵敏度为64.0%。联合2种方法平行检测dsDNA抗体可明显提高SLE的诊断效能, ROC曲线下面积最高（0.869 0）。结论：单独应用ELISA法时, 应警惕假阳性较高, 需排除其他非SLE疾病的诊断;联合使用ELISA法和CLIFT法检测抗dsDNA抗体可较单独应用CLIFT法明显提高SLE的诊断效能, 二者联合平行检测结果为阴性, 用于排除SLE更有意义。
Objective: To analyze the occurrence of anti-double-stranded DNA (dsDNA) IgG antibody in SLE (systemic lupus erythematosus) and non-SLE patients and the efficacy of combined use of ELISA and CLIFT (crithidia luciliae immunofluorescence test) in detection of anti-dsDNA IgG antibody for diagnosis of SLE. Methods: The clinical data of 4 726 patients undergoing anti-dsDNA IgG antibody detection by ELISA and 830 of them receiving CLIFT were collected. Receiver operating characteristic curve (ROC) was used to evaluate the performance of each or combined use of the two in detection of anti-dsDNA IgG antibody for the diagnosing of SLE. Results: Among the 4 726 patients tested by ELISA, 398 cases were anti-dsDNA IgG antibody positive. Of them 183 patients were diagnosed as SLE (46.0%). The non-SLE patients included liver disease (28.4%), other connective tissue diseases (7.5%), hematological disease(5.3%) and respiratory di-sease(3.8%). For 830 cases undergoing CLIFT, 134 cases were anti-dsDNA IgG antibody positive, and 94% of them were diagnosed as SLE; the non-SLE patients included 3 cases of autoimmune liver disease, 1 case of liver cirrhosis, 1 case of nephritis, 1 case of psoriasis and 1 case of anti-phospholipid syndrome. In the cohort of 830 patient with both detection of anti-dsDNA IgG antibody by both ELISA and CLIFT, 197 patients were diagnosed as SLE, and 633 patients diagnosed as non-SLF. ELISA had a sensitivity of 74.6% and a specificity of 92.9%, while CLIFT had a specificity of 98.7% and a sensitivity of 64%. The combined detection by ELISA and CLIFT could significantly improve the efficacy of diagnosing SLE, with the highest AUC of 0.8 690. Conclusions: When ELISA is applied alone for detection of anti-dsDNA IgG antibody,a high false positivity should be paid attention to. The detection of anti-dsDNA IgG antibody by both ELISA and CLIFT could significantly improve the diagnostic efficacy for SLE than by CLIFT alone.
[1] Compagno M, Rekvig OP, Bengtsson AA, et al.Clinical phenotype associations with various types of anti-dsDNA antibodies in patients with recent onset of rheumatic symptoms. Results from a multicentre observational study[J]. Lupus Sci Med,2014,1(1):e000007.
[2] Enocsson H, Sjöwall C, Wirestam L, et al.Four Anti-dsDNA Antibody Assays in Relation to Systemic Lupus Erythematosus Disease Specificity and Activity[J]. J Rheumatol,2015,42(5):817-825.
[3] Almeida González D, Roces Varela A, Marcelino Rodríguez I, et al.Anti-dsDNA antibodies in systemic lupus erythematosus: A combination of two quantitative metho-ds and the ANA pattern is the most efficient strategy of detection[J]. J Immunol Methods,2015,427:30-35.
[4] Piga M, Floris A, Mathieu A, et al.Comment on: Auto-antibodies to double-stranded DNA as biomarker in systemic lupus erythematosus: comparison of different assays during quiescent and active disease[J]. Rheumatology (Oxford),2017,56(5):2038-2039.
[5] Infantino M, Meacci F, Bentow C, et al.Clinical compa-rison of QUANTA Flash dsDNA chemiluminescent immunoassay with four current assays for the detection of anti-dsDNA autoantibodies[J]. J Immunol Res,2015,2015:902821.
[6] Lakos G, Gonzalez M, Flaherty D, et al.Detection of anti-dsDNA antibodies by computer-aided automated immunofluorescence analysis[J]. J Immunol Methods,2016, 433:17-22.
[7] Mahler M, Bentow C, O'Malley T, et al. Performance Characteristics of Different Anti-Double-Stranded DNA Antibody Assays in the Monitoring of Systemic Lupus Erythematosus[J]. J Immunol Res,2017,2017:1720902.
[8] Compagno M, Jacobsen S, Rekvig OP, et al.Low diagnostic and predictive value of anti-dsDNA antibodies in unselected patients with recent onset of rheumatic symptoms: results from a long-term follow-up Scandinavian multicentre study[J]. Scand J Rheumatol,2013,42(4):311-316.
[9] 陈顺乐, 沈南, 顾越英, 等.系统性红斑狼疮的发病机理及诊治策略[Z]. 上海交通大学医学院附属仁济医院. 2007.
[10] 王慧芬, 李进. 肝病诊疗指南[M]. 北京: 军事医学出版社,2012.
[11] 张之南, 沈悌. 血液病诊断及疗效标准[M]. 北京: 科学出版社,1999.
[12] Viriyataveekul R, Kobkitjaroen J, Jaiyen J, et al.Evaluation of five commercial assays for the detection of anti-dsDNA antibodies: three Crithidia luciliae indirect immunofluorescence test kits and two enzyme immunoassay kits[J]. J Med Assoc Thai,2014,97(2):220-224.
[13] Novak GV, Molinari BC, Ferreira JC, et al.Characteristics of 1555 childhood-onset lupus in three groups based on distinct time intervals to disease diagnosis: a Brazilian multicenter study[J]. Lupus,2018,27(10):1712-1717.
[14] Poulsen NN, Pedersen ME, Østergaard J, et al.Flow-Induced Dispersion Analysis for Probing Anti-dsDNA Antibody Binding Heterogeneity in Systemic Lupus Erythematosus Patients: Toward a New Approach for Diagnosis and Patient Stratification[J]. Anal Chem,2016,88(18):9056-9061.
[15] Ru JL, Zhao Y, Xie XX, et al.Clinical applications of the indirect immunofluorescence assay for detection of anticell membrane-associated DNA antibodies in juvenile systemic lupus erythematosus[J]. Pediatr Res,2015,77(2):376-380.
[16] Sarbu MI, Salman-Monte TC, Rubio Muñoz P, et al.Differences between clinical and laboratory findings in patients with recent diagnosis of SLE according to the positivity of anti-dsDNA by the Crithidia luciliae method[J]. Lupus,2015,24(11):1198-1203.
[17] Hormann W, Hahn M, Gerlach S, et al.Performance analysis of automated evaluation of Crithidia luciliae-based indirect immunofluorescence tests in a routine setting-strengths and weaknesses[J]. Clin Chem Lab Med,2017,56(1):86-93.
[18] Monzavi SM, Alirezaei A, Shariati-Sarabi Z, et al.Efficacy analysis of hydroxychloroquine therapy in systemic lupus erythematosus: a study on disease activity and immunological biomarkers[J]. Inflammopharmacology,2018.
[19] Mosca M, Costenbader KH, Johnson SR, et al.How Do Patients With Newly Diagnosed Systemic Lupus Erythematosus Present? A Multicenter Cohort of Early Systemic Lupus Erythematosus to Inform the Development of New Classification Criteria[J]. Arthritis Rheumatol,2018.
[20] Yang J, Xu Z, Sui M, et al.Co-Positivity for Anti-dsDNA, -Nucleosome and -Histone Antibodies in Lupus Nephritis Is Indicative of High Serum Levels and Severe Nephropathy[J]. PLoS One,2015,10(10):e0140441.
[21] Yeganeh H, Parvaneh j, Mehregan F, et al. Autoimmune Hepatitis as an Initial Presentation of SLE[J]. Arch Pe-diatr Infect Dis,2016:e34653.
