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Comparison study of bead-based and line-blot multiplex ANA immunoassays in the diagnosis of systemic autoimmune rheumatic diseases

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Abstract

Objectives

Detection of antinuclear antibodies (ANA) by immunofluorescence assay (IFA) is increasingly substituted by multiplex bead-based immunoassay (MBA) and line-blot immunoassay (LIA). This study is to compare the diagnostic performance of MBA and LIA ANA assays on clinically characterized patient samples.

Methods

A total of 728 serum samples from 385 patients with systemic autoimmune rheumatic diseases (SARD), 204 patients with non-SARD diseases, and 139 apparently healthy subjects were tested with the BioPlex 2200 ANA Screen and EuroLine ANA Profile 3 as the representative MBA and LIA technologies and HEp-2 ANA IFA. Clinical data were collected independent of laboratory analysis and later related to the ANA test results. The clinical diagnostic performances were analyzed using Analyse-it software.

Results

The MBA demonstrated higher area under curve (AUC) compared to LIA (0.814 vs 0.761, p = 0.002) and HEp-2 IFA (0.814 vs 0.771, p = 0.008). The MBA and LIA ANA methods showed higher specificity (83.8% and 77.0% vs 67.6%, p < 0.001 and p = 0.005) but lower sensitivity (79.0% and 75.3% vs 86.5%, p < 0.001) compared to HEp-2 IFA. The MBA and LIA ANA revealed substantial to excellent agreements on specific antinuclear antibodies except anti-dsDNA, with the total agreement from 92.3 to 99.9% and Cohen’s kappa from 0.71 to 0.98. The MBA demonstrated significantly higher sensitivity (58.1% vs 19.8%, p < 0.001) and comparable specificity (95.9% vs 97.5%, p = 0.221) on anti-dsDNA assay for the diagnosis of SLE compared to LIA.

Conclusions

The MBA and LIA ANA assays have higher specificity but lower sensitivity compared to HEp-2 IFA. There are good agreements between MBA and LIA ANA for the specific antinuclear antibodies except for anti-dsDNA. The MBA ANA demonstrated better assay performance compared to LIA as the MBA possesses higher sensitivity and specificity in the diagnosis of SARD.

Key Points

• The multiplex bead-based immunoassay (MBA) ANA outperformed line-blot immunoassay (LIA) and traditional HEp-2 IFA.

• There are good agreements between the MBA BioPlex 2200 ANA Screen and LIA EuroLine ANA Profile 3 for the most of specific antinuclear antibodies except anti-dsDNA.

• Additional anti-dsDNA testing is suggested when EuroLine ANA Profile 3 is used for the aid of SLE diagnosis and management.

• The positive predictive value of both multiplex ANA assays can be substantially increased without significantly affecting negative predictive value by using at least two specific antinuclear antibodies for reporting a positive ANA result.

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Data availability

The datasets generated and/or analyzed during the current study will be made available in DataverseNL after publication. They are also available from the corresponding author on request.

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Funding

This work was supported by the National Key Clinical Specialty (grant number: 2012649).

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Authors and Affiliations

  1. Department of Dermatology and Rheumatology, Rui** Hospital, Shanghai Jiao Tong University School of Medicine, 197 Rui** Er Road, Shanghai, 200025, China

    Weiru Yuan, Hua Cao & Jie Zheng

  2. Laboratory of Dermatoimmunology, Rui** Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China

    Wei** Li & **nyi Wu

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  1. Weiru Yuan
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  2. Hua Cao
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  3. Wei** Li
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Contributions

JZ and WY contributed to the conception and design of the study. WY, HC, WL, and XW conducted the experiments and data collection. WY and JZ analyzed the data and wrote the manuscript. All authors have approved the final version of the manuscript to be submitted.

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Correspondence to Jie Zheng.

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Informed consents were obtained from all participants in the study, and the study protocol was approved by the Rui** Hospital Ethics Committee of Shanghai Jiao Tong University, School of Medicine, Shanghai, China, in compliance with the principles of the Helsinki Declaration.

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Yuan, W., Cao, H., Li, W. et al. Comparison study of bead-based and line-blot multiplex ANA immunoassays in the diagnosis of systemic autoimmune rheumatic diseases. Clin Rheumatol 41, 899–909 (2022). https://doi.org/10.1007/s10067-021-05946-7

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