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Peer-reviewedCC BY 4.0PLOS One

ANCA testing strategies: A single centre experience over the past decade in a large teaching hospital in the Netherlands

8 affiliations
  1. 1Department of Pathology, Leiden University Medical Center, Leiden, The Netherlands
  2. 2Pathology Laboratory, Pathan B.V., Rotterdam, The Netherlands
  3. 3Department of Rheumatology and Clinical Immunology, Maasstad Hospital, Rotterdam, The Netherlands
  4. 4Business Intelligence, Maasstad Hospital, Rotterdam, The Netherlands
  5. 5Department of Pathology and Medical Biology, University Medical Center Groningen, Groningen, The Netherlands
  6. 6Department of Internal Medicine, Maasstad Hospital, Rotterdam, The Netherlands
  7. 7Laboratory of Clinical Chemistry, MaasstadLab, Maasstad Hospital, Rotterdam, The Netherlands
  8. 8Department of Rheumatology, Leiden University Medical Center, Leiden, The Netherlands

Abstract

Objectives: In 2017 a new consensus for antineutrophil cytoplasmic antibody (ANCA) testing was published, based on data from tertiary referral centres for ANCA-associated vasculitis (AAV). AAV-prevalence in their tested population was relatively high. We analysed how the consensus approach performs in a large teaching hospital and tailored it to secondary care settings with lower patient prevalences in their population. Methods: Patients screened for ANCA between 2012–2023 were included; clinical data were retrospectively collected. Indirect immunofluorescence (IIF) was performed using EUROPLUSTM Granulocyte Mosaic 25 IIF. MPO/PR3 was detected by ImmunoCAP®250. Statistical analyses were performed in R. Results: 286/5518 (5.2%) patients tested for ANCA were positive: 63% had no AAV. Non-AAV patients often had other autoimmune diseases, malignancies, infections or used specific drugs. IIF and ELISA as first test showed good negative predictive values. ELISA as first test showed higher positive predictive value and specificity; performing a second test increased specificity. ANCA concentrations were higher in AAV than non-AAV, but ranges were wide. Conclusion: The majority of ANCA-positive patients did not have AAV. In our secondary care setting, ELISA performs best as first test to rule out and to rule in AAV. A second test should be considered with high clinical suspicion and a negative first test, or with low antibody concentrations, following the consensus. Additionally, we propose a second test when there is clinical doubt and a positive first test. This increases specificity and PPV, reducing false positives, which is especially relevant in secondary care settings.

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BibTeX
@article{trejo2026anca,
  title = {{ANCA testing strategies: A single centre experience over the past decade in a large teaching hospital in the Netherlands}},
  author = {Maria A. C. Wester Trejo and Tjallingius Martijn Kuijper and Geeke J. Waverijn and Ingeborg M. Bajema and René M. A. van den Dorpel and Marc R. Kok and Iris J. A. M. Verberk-Jonkers and Els J. M. Zirkzee and Snjezana Kos and Annelies E. Berden},
  journal = {PLOS One},
  year = {2026},
  month = sep,
  doi = {10.1371/journal.pone.0357495},
  url = {https://doi.org/10.1371/journal.pone.0357495},
  note = {CC BY 4.0},
}
RIS
TY  - JOUR
TI  - ANCA testing strategies: A single centre experience over the past decade in a large teaching hospital in the Netherlands
AU  - Maria A. C. Wester Trejo
AU  - Tjallingius Martijn Kuijper
AU  - Geeke J. Waverijn
AU  - Ingeborg M. Bajema
AU  - René M. A. van den Dorpel
AU  - Marc R. Kok
AU  - Iris J. A. M. Verberk-Jonkers
AU  - Els J. M. Zirkzee
AU  - Snjezana Kos
AU  - Annelies E. Berden
PY  - 2026
DA  - 2026/09/09
JO  - PLOS One
DO  - 10.1371/journal.pone.0357495
UR  - https://doi.org/10.1371/journal.pone.0357495
AB  - Objectives: In 2017 a new consensus for antineutrophil cytoplasmic antibody (ANCA) testing was published, based on data from tertiary referral centres for ANCA-associated vasculitis (AAV). AAV-prevalence in their tested population was relatively high. We analysed how the consensus approach performs in a large teaching hospital and tailored it to secondary care settings with lower patient prevalences in their population. Methods: Patients screened for ANCA between 2012–2023 were included; clinical data were retrospectively collected. Indirect immunofluorescence (IIF) was performed using EUROPLUSTM Granulocyte Mosaic 25 IIF. MPO/PR3 was detected by ImmunoCAP®250. Statistical analyses were performed in R. Results: 286/5518 (5.2%) patients tested for ANCA were positive: 63% had no AAV. Non-AAV patients often had other autoimmune diseases, malignancies, infections or used specific drugs. IIF and ELISA as first test showed good negative predictive values. ELISA as first test showed higher positive predictive value and specificity; performing a second test increased specificity. ANCA concentrations were higher in AAV than non-AAV, but ranges were wide. Conclusion: The majority of ANCA-positive patients did not have AAV. In our secondary care setting, ELISA performs best as first test to rule out and to rule in AAV. A second test should be considered with high clinical suspicion and a negative first test, or with low antibody concentrations, following the consensus. Additionally, we propose a second test when there is clinical doubt and a positive first test. This increases specificity and PPV, reducing false positives, which is especially relevant in secondary care settings.
SN  - 1932-6203
ER  -