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Detection of antineutrophil cytoplasmic antibodies (ANCAs): a multicentre European Vasculitis Study Group (EUVAS) evaluation of the value of indirect immunofluorescence (IIF) versus antigen-specific immunoassays
  1. Jan Damoiseaux1,
  2. Elena Csernok2,
  3. Niels Rasmussen3,
  4. Frank Moosig4,
  5. Pieter van Paassen5,
  6. Bo Baslund6,
  7. Pieter Vermeersch7,8,
  8. Daniel Blockmans9,
  9. Jan-Willem Cohen Tervaert10,
  10. Xavier Bossuyt11,12
  1. 1Central Diagnostic Laboratory, Maastricht University Medical Center, Maastricht, The Netherlands
  2. 2Department of Rheumatology and Immunology, Klinikum Bad Bramstedt, Bad Bramstedt, Germany
  3. 3Department of Autoimmune Serology, Statens Seruminstitute, Copenhagen, Denmark
  4. 4Rheumazentrum Schleswig-Holstein Mitte, Neumünster, Germany
  5. 5Department of Internal Medicine, Section Nephrology and Immunology, Maastricht University Medical Center, Maastricht, The Netherlands
  6. 6Department of Rheumatology, Rigshospitalet, Copenhagen, Denmark
  7. 7Clinical Department of Laboratory Medicine, University Hospitals Leuven, Leuven, Belgium
  8. 8Department of Cardiovascular Sciences, KU Leuven, Leuven, Belgium
  9. 9Clinical Department of General Internal Medicine, Research Department of Microbiology and Immunology, Laboratory of Clinical Infectious and Inflammatory Disorders, University Hospitals Leuven, Leuven, Belgium
  10. 10Maastricht University, Maastricht, The Netherlands
  11. 11Department of Microbiology and Immunology, KU Leuven, Leuven, Belgium
  12. 12Department of Laboratory Medicine, University Hospitals Leuven, Leuven, Belgium
  1. Correspondence to Dr Xavier Bossuyt, Laboratory Medicine, University Hospitals Gasthuisberg, Herestraat 49, Leuven 3000, Belgium; Xavier.Bossuyt{at}


Objective This multicentre study was performed to evaluate the diagnostic accuracy of a wide spectrum of novel technologies nowadays available for detection of myeloperoxidase (MPO) and proteinase 3 (PR3)-antineutrophil cytoplasmic antibodies (ANCAs).

Methods Sera (obtained at the time of diagnosis) from 251 patients with ANCA-associated vasculitis (AAV), including granulomatosis with polyangiitis and microscopic polyangiitis, and from 924 disease controls were tested for the presence of cytoplasmic pattern/perinuclear pattern and atypical ANCA (A-ANCA) by indirect immunofluorescence (IIF) (at two sites) and for the presence of PR3-ANCA and MPO-ANCA by eight different immunoassays.

Results The area under the curve (AUC) of the receiver operating characteristic curve to discriminate AAV from controls was 0.923 (95% CI 0.902 to 0.944) and 0.843 (95% CI 0.814 to 0.871) for the two IIF methods. For the antigen-specific immunoassays, the AUC varied between 0.936 (95% CI 0.912 to 0.960) and 0.959 (95% CI 0.941 to 0.976), except for one immunoassay for which the AUC was 0.919 (95% CI 0.892 to 0.945).

Conclusions Our comparison of various ANCA detection methods showed (i) large variability between the two IIF methods tested and (ii) a high diagnostic performance of PR3-ANCA and MPO-ANCA by immunoassay to discriminate AAV from disease controls. Consequently, dual IIF/antigen-specific immunoassay testing of each sample is not necessary for maximal diagnostic accuracy. These results indicate that the current international consensus on ANCA testing for AAV needs revision.

  • Systemic vasculitis
  • Autoantibodies
  • Autoimmune Diseases

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