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Staining and serological characteristics of the AC-30 pattern in HEp-2 IIFA: a comparative study with AC-2 pattern

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Published/Copyright: September 25, 2025
Clinical Chemistry and Laboratory Medicine (CCLM)
From the journal Clinical Chemistry and Laboratory Medicine (CCLM) Volume 64 Issue 2

Abstract

Objectives

To investigate the staining and serological characteristics of AC-30 pattern.

Methods

A total of 184 participants were recruited from patients who underwent routine antinuclear antibody testing between 2022 and 2023 at Peking Union Medical College Hospital. Cohort 1 (n=47) showed the AC-30 pattern on HEp-2 indirect immunofluorescence assay, and cohort 2 (n=137) showed AC-2 pattern as control. Anti-DFS70 antibody detection and DFS70 antigen adsorption assays were conducted. Pattern simulation assays were performed by combining serum samples exhibiting classic AC-2 pattern with other common HEp-2 IIFA patterns.

Results

Anti-DFS70 antibodies were detected in 97 % of patients in cohort one and in all patients in cohort 2. The titers of the IIFA pattern showed a weak correlation with anti-DFS70 antibody levels in cohort 1 (r=0.35, p=0.0331). In DFS70 antigen adsorption assays, a higher proportion of homogeneous nuclear staining was observed in cohort 1 (79 %) than in cohort 2 (62 %) (p=0.037). Simulated samples mixed classic AC-2 with homogeneous pattern resembled those of AC-30 pattern in both interphase and mitotic cells. Especially, the staining characteristics of AC-2 became increasingly indistinct when mixed with higher-titer homogeneous patterns. Among samples exhibiting homogeneous patterns post-DFS70 immunoadsorption, non-autoimmune conditions were more common in cohort one than cohort two.

Conclusions

The presence of relatively lower anti-DFS70 antibodies levels or coexisting high-titer homogeneous patterns may contribute to the development of the AC-30 pattern rather than AC-2. This finding needs to be further confirmed in larger-scale studies.

Keywords: AC-30; AC-2; DFS; anti-DFS70 antibody; HEp-2 indirect immunofluorescence assay
Corresponding authors: Chaojun Hu, Department of Rheumatology and Clinical Immunology, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, China; National Clinical Research Center for Dermatologic and Immunologic Diseases (NCRC-DID), Ministry of Science & Technology, Beijing, China; State Key Laboratory of Complex Severe and Rare Diseases, Peking Union Medical College Hospital, Beijing, China; Key Laboratory of Rheumatology and Clinical Immunology, Ministry of Education, Beijing, China; and Shuaifuyuan, Dongcheng District, Beijing, 100730, China, E-mail: ; Yunyun Fei, Department of Rheumatology and Clinical Immunology, Peking Union Medical College Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, China; National Clinical Research Center for Dermatologic and Immunologic Diseases (NCRC-DID), Ministry of Science & Technology, Beijing, China; State Key Laboratory of Complex Severe and Rare Diseases, Peking Union Medical College Hospital, Beijing, China; Key Laboratory of Rheumatology and Clinical Immunology, Ministry of Education, Beijing, China; Department of Health Medicine, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing, China; and Shuaifuyuan, Dongcheng District, Beijing, 100730, China, E-mail: ; and Renfang Zhou, Department of Clinical Laboratory, The First People’s Hospital of Wenling, Affiliated to Wenzhou Medical University, Chuanan South Road, Chengxi Street, Wenling 317500, Taizhou, China, E-mail:
Chuiwen Deng, Ningxin Li and Ruxi Hu contributed equally to this work and share first authorship. Chaojun Hu, Yunyun Fei and Renfang Zhou share senior authorship.

Funding source: the National High-Level Hospital Clinical Research Funding

Award Identifier / Grant number: 2022-PUMCH-A-040

Funding source: the Medical Scientific Research Foundation of Zhejiang Province

Award Identifier / Grant number: 2022KY1413

  1. Research ethics: This study was approved by the Ethics Committee of Peking Union Medical College Hospital (No. I-22PJ930) and was conducted in accordance with the Declaration of Helsinki.

  2. Informed consent: Not applicable.

  3. Author contributions: All authors have accepted responsibility for the entire content of this manuscript and approved its submission.

  4. Use of Large Language Models, AI and Machine Learning Tools: None declared.

  5. Conflict of interest: The authors state no conflict of interest.

  6. Research funding: This study was support by the National High-Level Hospital Clinical Research Funding (2022-PUMCH-A-040); the Medical Scientific Research Foundation of Zhejiang Province (2022KY1413).

  7. Data availability: The data are available from the corresponding author on reasonable request.

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Supplementary Material

This article contains supplementary material (https://doi.org/10.1515/cclm-2025-0817).

Received: 2025-07-02
Accepted: 2025-09-07
Published Online: 2025-09-25
Published in Print: 2026-01-29

© 2025 Walter de Gruyter GmbH, Berlin/Boston

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https://doi.org/10.1515/cclm-2025-0817
Keywords for this article
AC-30; AC-2; DFS; anti-DFS70 antibody; HEp-2 indirect immunofluorescence assay

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  2. Editorials
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  4. Multi-cancer early detection revisited: insights and lessons from the PATHFINDER 2 study
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  9. Value is equal to outcome/costs: how to apply to laboratory medicine
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  12. Predictive genomic medicine enlarges the spectrum of predisposing mutations for head and neck cancers via a panel of 56 genes selected for human neoplasia in Southern Italy: a pilot study
  13. Failure Mode and Effects Analysis (FMEA) integrating quality indicators for risk assessment of the total testing process in human papillomavirus genotyping testing: a proactive risk analysis model for molecular diagnostics
  14. Impact of pneumatic tube system transport and blood collection tubes on cell-free DNA analysis of plasma samples
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  16. Requiem for the Passing–Bablok nonparametric regression in assessing the agreement between two measurement methods
  17. Comparative analysis of ISO 15189:2022 with earlier versions with reference to ethics
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