Comparative evaluation of three anti-dsDNA antibody detection methods in systemic lupus erythematosus: insights from a large monocentric cohort

BackgroundAnti-double-stranded DNA (anti-dsDNA) antibodies at abnormal titer are of considerable diagnostic value for systemic lupus erythematosus (SLE). Current assays detecting anti-dsDNA antibodies show divergent properties, emphasizing the importance of selecting suitable assays. This study aims...

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Main Authors: Ruijing Lu, Rui Yu, Rong Huang, Chiyuan Xue, Ning Song, Jiuliang Zhao, Xiaofeng Zeng, Chaojun Hu
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-04-01
Series:Frontiers in Immunology
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Online Access:https://www.frontiersin.org/articles/10.3389/fimmu.2025.1529484/full
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author Ruijing Lu
Ruijing Lu
Rui Yu
Rui Yu
Rong Huang
Rong Huang
Chiyuan Xue
Ning Song
Jiuliang Zhao
Xiaofeng Zeng
Chaojun Hu
author_facet Ruijing Lu
Ruijing Lu
Rui Yu
Rui Yu
Rong Huang
Rong Huang
Chiyuan Xue
Ning Song
Jiuliang Zhao
Xiaofeng Zeng
Chaojun Hu
author_sort Ruijing Lu
collection DOAJ
description BackgroundAnti-double-stranded DNA (anti-dsDNA) antibodies at abnormal titer are of considerable diagnostic value for systemic lupus erythematosus (SLE). Current assays detecting anti-dsDNA antibodies show divergent properties, emphasizing the importance of selecting suitable assays. This study aims to investigate the diagnostic performance of indirect immunofluorescence (IIF), digital liquid chip method (DLCM), chemiluminescence immunoassay (CLIA), and their combinations for detecting anti-dsDNA antibodies in SLE.MethodsWe conducted a retrospective, single-center study from 2022 to 2023 which included 3429 samples: 1773 from patients with SLE and 1656 from controls with rheumatoid arthritis (RA) and Sjögren’s syndrome (SS). Sensitivity, specificity, accuracy, positive predictive value (PPV), and negative predictive value (NPV) for anti-dsDNA detection by IIF, DLCM, and CLIA were calculated. Cohen’s kappa coefficient was used to evaluate inter-method agreement. The correlations between anti-dsDNA concentration and SLEDAI-2k scores/renal involvement were assessed.ResultsAmong individual assays, IIF demonstrated the highest specificity (98.31%) and PPV (96.10%) but lower sensitivity (38.92%) compared to CLIA (41.57%) and DLCM (43.65%) (p < 0.05). Combining two assays significantly improved sensitivity while maintaining specificity>95%. The combination of IIF and DLCM achieved a sensitivity of 52.2% and an AUC of 0.76. Substantial agreement was observed between DLCM and CLIA (κ = 0.78), whereas agreement between IIF and the other assays was moderate (κ = 0.65–0.66). In a longitudinal analysis of 88 SLE patients, CLIA and DLCM detected antibody fluctuations more reliably than IIF. Anti-dsDNA levels by DLCM or CLIA positively correlated with SLEDAI-2K scores (R=0.42 and 0.29, p<0.05). Both IIF and CLIA methods showed significant differences between the SLE patients with and without renal involvement (p < 0.05). The combination of two assays provided higher sensitivity than single assays (p<0.001) in renal involvement subgroups.ConclusionOur findings demonstrate that DLCM performs comparably to CLIA, supporting its clinical potential. Moreover, combining assays significantly enhances diagnostic sensitivity, particularly in subgroups with renal involvement.
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spelling doaj-art-b0359ff152fc452293d45297412f3be22025-08-20T03:28:58ZengFrontiers Media S.A.Frontiers in Immunology1664-32242025-04-011610.3389/fimmu.2025.15294841529484Comparative evaluation of three anti-dsDNA antibody detection methods in systemic lupus erythematosus: insights from a large monocentric cohortRuijing Lu0Ruijing Lu1Rui Yu2Rui Yu3Rong Huang4Rong Huang5Chiyuan Xue6Ning Song7Jiuliang Zhao8Xiaofeng Zeng9Chaojun Hu10Department of Rheumatology and Clinical Immunology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Peking Union Medical College, National Clinical Research Center for Dermatologic and Immunologic Diseases (NCRC-DID), Ministry of Science & Technology, Key Laboratory of Rheumatology and Clinical Immunology, Ministry of Education, Beijing, ChinaDepartment of Laboratory Medicine, Shenzhen Baoan Women’s and Children’s Hospital, Shenzhen, ChinaDepartment of Rheumatology and Clinical Immunology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Peking Union Medical College, National Clinical Research Center for Dermatologic and Immunologic Diseases (NCRC-DID), Ministry of Science & Technology, Key Laboratory of Rheumatology and Clinical Immunology, Ministry of Education, Beijing, ChinaEight-year Medical Doctor Program, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, ChinaDepartment of Rheumatology and Clinical Immunology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Peking Union Medical College, National Clinical Research Center for Dermatologic and Immunologic Diseases (NCRC-DID), Ministry of Science & Technology, Key Laboratory of Rheumatology and Clinical Immunology, Ministry of Education, Beijing, ChinaDepartment of Clinical Laboratory, The People’s Hospital of Yuhuan, Taizhou, Zhejiang, ChinaDepartment of Rheumatology and Clinical Immunology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Peking Union Medical College, National Clinical Research Center for Dermatologic and Immunologic Diseases (NCRC-DID), Ministry of Science & Technology, Key Laboratory of Rheumatology and Clinical Immunology, Ministry of Education, Beijing, ChinaDepartment of Rheumatology and Clinical Immunology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Peking Union Medical College, National Clinical Research Center for Dermatologic and Immunologic Diseases (NCRC-DID), Ministry of Science & Technology, Key Laboratory of Rheumatology and Clinical Immunology, Ministry of Education, Beijing, ChinaDepartment of Rheumatology and Clinical Immunology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Peking Union Medical College, National Clinical Research Center for Dermatologic and Immunologic Diseases (NCRC-DID), Ministry of Science & Technology, Key Laboratory of Rheumatology and Clinical Immunology, Ministry of Education, Beijing, ChinaDepartment of Rheumatology and Clinical Immunology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Peking Union Medical College, National Clinical Research Center for Dermatologic and Immunologic Diseases (NCRC-DID), Ministry of Science & Technology, Key Laboratory of Rheumatology and Clinical Immunology, Ministry of Education, Beijing, ChinaDepartment of Rheumatology and Clinical Immunology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Peking Union Medical College, National Clinical Research Center for Dermatologic and Immunologic Diseases (NCRC-DID), Ministry of Science & Technology, Key Laboratory of Rheumatology and Clinical Immunology, Ministry of Education, Beijing, ChinaBackgroundAnti-double-stranded DNA (anti-dsDNA) antibodies at abnormal titer are of considerable diagnostic value for systemic lupus erythematosus (SLE). Current assays detecting anti-dsDNA antibodies show divergent properties, emphasizing the importance of selecting suitable assays. This study aims to investigate the diagnostic performance of indirect immunofluorescence (IIF), digital liquid chip method (DLCM), chemiluminescence immunoassay (CLIA), and their combinations for detecting anti-dsDNA antibodies in SLE.MethodsWe conducted a retrospective, single-center study from 2022 to 2023 which included 3429 samples: 1773 from patients with SLE and 1656 from controls with rheumatoid arthritis (RA) and Sjögren’s syndrome (SS). Sensitivity, specificity, accuracy, positive predictive value (PPV), and negative predictive value (NPV) for anti-dsDNA detection by IIF, DLCM, and CLIA were calculated. Cohen’s kappa coefficient was used to evaluate inter-method agreement. The correlations between anti-dsDNA concentration and SLEDAI-2k scores/renal involvement were assessed.ResultsAmong individual assays, IIF demonstrated the highest specificity (98.31%) and PPV (96.10%) but lower sensitivity (38.92%) compared to CLIA (41.57%) and DLCM (43.65%) (p < 0.05). Combining two assays significantly improved sensitivity while maintaining specificity>95%. The combination of IIF and DLCM achieved a sensitivity of 52.2% and an AUC of 0.76. Substantial agreement was observed between DLCM and CLIA (κ = 0.78), whereas agreement between IIF and the other assays was moderate (κ = 0.65–0.66). In a longitudinal analysis of 88 SLE patients, CLIA and DLCM detected antibody fluctuations more reliably than IIF. Anti-dsDNA levels by DLCM or CLIA positively correlated with SLEDAI-2K scores (R=0.42 and 0.29, p<0.05). Both IIF and CLIA methods showed significant differences between the SLE patients with and without renal involvement (p < 0.05). The combination of two assays provided higher sensitivity than single assays (p<0.001) in renal involvement subgroups.ConclusionOur findings demonstrate that DLCM performs comparably to CLIA, supporting its clinical potential. Moreover, combining assays significantly enhances diagnostic sensitivity, particularly in subgroups with renal involvement.https://www.frontiersin.org/articles/10.3389/fimmu.2025.1529484/fullanti-double-stranded DNAsystemic lupus erythematosusindirect immunofluorescence assaydigital liquid chip methodchemiluminescence immunoassay
spellingShingle Ruijing Lu
Ruijing Lu
Rui Yu
Rui Yu
Rong Huang
Rong Huang
Chiyuan Xue
Ning Song
Jiuliang Zhao
Xiaofeng Zeng
Chaojun Hu
Comparative evaluation of three anti-dsDNA antibody detection methods in systemic lupus erythematosus: insights from a large monocentric cohort
Frontiers in Immunology
anti-double-stranded DNA
systemic lupus erythematosus
indirect immunofluorescence assay
digital liquid chip method
chemiluminescence immunoassay
title Comparative evaluation of three anti-dsDNA antibody detection methods in systemic lupus erythematosus: insights from a large monocentric cohort
title_full Comparative evaluation of three anti-dsDNA antibody detection methods in systemic lupus erythematosus: insights from a large monocentric cohort
title_fullStr Comparative evaluation of three anti-dsDNA antibody detection methods in systemic lupus erythematosus: insights from a large monocentric cohort
title_full_unstemmed Comparative evaluation of three anti-dsDNA antibody detection methods in systemic lupus erythematosus: insights from a large monocentric cohort
title_short Comparative evaluation of three anti-dsDNA antibody detection methods in systemic lupus erythematosus: insights from a large monocentric cohort
title_sort comparative evaluation of three anti dsdna antibody detection methods in systemic lupus erythematosus insights from a large monocentric cohort
topic anti-double-stranded DNA
systemic lupus erythematosus
indirect immunofluorescence assay
digital liquid chip method
chemiluminescence immunoassay
url https://www.frontiersin.org/articles/10.3389/fimmu.2025.1529484/full
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