Comprehensive evaluation of antiphospholipid antibody testing methodologies in APS diagnosis: performance comparisons across assay systems and clinical subtypes
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Yun Wang
,
Hongyan Hou
und
Shiji Wu
Abstract
Objectives
Antiphospholipid syndrome (APS) is an autoimmune disorder characterized by thrombosis and obstetric complications associated with antiphospholipid antibodies (aPLs). This study aimed to compare the diagnostic performance of six commercial assay systems for detecting aCL and aβ2GPI antibodies.
Methods
Sixty-three APS patients, 50 SLE patients, 67 disease controls, and 62 healthy controls were enrolled. aCL and aβ2GPI antibodies of IgA, IgG, and IgM isotypes were measured using six commercial platforms, including three ELISA-based systems and three CLIA-based systems. Inter-assay concordance was compared across all detection platforms, and ROC curve analysis was performed to evaluate and compare their diagnostic performance in APS.
Results
Inter-assay concordance varied across platforms, with IgG isotypes showing the highest consistency and IgA exhibiting the lowest agreement. Overall, CLIA-based systems demonstrated superior classification performance compared to ELISA-based methods. The highest area under the curve (AUC) reached 0.811, with sensitivity and specificity up to 0.730 and 0.891, respectively. IgG isotypes demonstrated the best overall performance, while IgA and IgM showed greater variability. The inclusion of IgA modestly improved sensitivity in some systems, although this was sometimes accompanied by decreased specificity. LA-positive patients had higher aPL positivity rates than LA-negative ones, and aPL levels were higher in thrombotic vs. obstetric APS.
Conclusions
Significant variability exists among commercial aPL detection systems. CLIA-based methods provided better consistency and diagnostic accuracy than ELISA. The inclusion of IgA provided additional diagnostic value in identifying APS patients who tested negative for aCL and aβ2GPI of the IgG and IgM isotypes.
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Research ethics: This study was approved by the ethical committee of Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology (TJ-IRB2023S204).
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Informed consent: Written informed consent to participate in this study was provided by the participants.
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Author contributions: YW and SW wrote the manuscript. XY, WW, RC collected the materials of patients. RO and TW did the assay detection. FW, HH and SW analyzed the data and helped revise the manuscript.
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Use of Large Language Models, AI and Machine Learning Tools: None declared.
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Conflict of interest: The authors state no conflict of interest.
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Research funding: This work was supported by grants from the Hubei Provincial Health Commission (WJ2023M010).
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Data availability: All data generated or analyzed during this study are included in this published article.
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Supplementary Material
This article contains supplementary material (https://doi.org/10.1515/cclm-2025-0499).
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