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Performance evaluation of a commercial line blot assay system for detection of myositis- and systemic sclerosis-related autoantibodies

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Abstract

Introduction/Objectives

A line blot (LB) assay is a multi-analyte platform capable of simultaneously detecting multiple anti-nuclear antibody specificities. Here, we evaluated the performance of a commercial LB assay developed for the identification of myositis- or systemic sclerosis (SSc)-related autoantibodies (autoAbs).

Method

We screened 300 serum samples from patients with various connective tissue diseases using an LB assay and compared the results of myositis- or SSc-related autoAbs with those identified by RNA and protein immunoprecipitation (IP) assays or indirect immunofluorescence (IIF).

Results

The IP assays revealed anti-Jo-1 Abs in 14 patients, anti-EJ Abs in 12, anti-PL-7 Abs in 8, anti-PL-12 Abs in 4, anti-Mi-2 Abs in 6, anti-SRP Abs in 8, anti-topoisomerase I Abs in 54, anti-RNA polymerase III Abs in 24, anti-U3 RNP Abs in 9, anti-Th/To Abs in 9, anti-Ku Abs in 14 and anti-hUBF Abs in 4, whereas IIF identified anti-centromere in 35. Good agreement between the IP assays and the LB assay was found only for anti-Jo-1 and anti-centromere antibodies. When a cut-off was adjusted to reconcile with the results of IP assays, the detection performance of LB assay was improved for anti-EJ, anti-PL-7, anti-PL-12, anti-SRP, anti-topoisomerase I and anti-RNA polymerase III Abs. However, the results of anti-Mi-2, anti-U3 RNP, anti-Th/To, anti-hUBF and anti-Ku Abs remained discordant between the LB assay and IP assays at all cut-off levels.

Conclusions

Detection of myositis- or SSc-related autoAbs using a commercial LB assay requires great caution since it can yield analytically false-positive or false-negative results.

Key Points

• A line blot (LB) assay is a multi-analyte platform capable of simultaneously detecting multiple antibodies with anti-nuclear specificities.

• Detection of myositis- or systemic sclerosis-related autoantibodies using a commercial LB assay requires great caution since it can yield analytically false-positive or false-negative results.

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Acknowledgments

We thank Ms. Masako Matsubara and Natsuho Yoshifuji for their technical assistance.

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Authors and Affiliations

  1. Department of Dermatology, Faculty of Medicine, Institute of Medical, Pharmaceutical and Health Sciences, Kanazawa University, 13-1 Takaramachi, Kanazawa, Ishikawa, 920-8641, Japan

    Yasuhito Hamaguchi & Kazuhiko Takehara

  2. Department of Allergy and Rheumatology, Nippon Medical School Graduate School of Medicine, Tokyo, Japan

    Masataka Kuwana

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  1. Yasuhito Hamaguchi
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  2. Masataka Kuwana
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  3. Kazuhiko Takehara
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Correspondence to Yasuhito Hamaguchi.

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All study participants provided written informed consent. The protocol was approved by Kanazawa University and Keio University.

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M.K. received patent fees from MBL, INOVA, and Phadia. Other authors have no conflict of interest to declare.

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Hamaguchi, Y., Kuwana, M. & Takehara, K. Performance evaluation of a commercial line blot assay system for detection of myositis- and systemic sclerosis-related autoantibodies. Clin Rheumatol 39, 3489–3497 (2020). https://doi.org/10.1007/s10067-020-04973-0

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  • DOI: https://doi.org/10.1007/s10067-020-04973-0

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