Detection of Anti-neutrophil Cytoplasmic Antibodies (ANCA) by Indirect Immunofluorescence

  • J. J. B. C. van Beers
  • J. Vanderlocht
  • C. Roozendaal
  • J. Damoiseaux
Part of the Methods in Molecular Biology book series (MIMB, volume 1901)


The eventual presence of anti-neutrophil cytoplasmic antibodies (ANCA) can initially be screened with indirect immunofluorescence (IIF). The majority of laboratories that facilitate ANCA testing use commercial kits. Although in-house assays are not encouraged in routine clinical laboratories, knowledge on the methodological aspects of the assay remains of importance. These aspects include choice of substrate, choice of fixative, staining procedure, and interpretation procedure. In this paper details on the methodology are provided and discussed in the context of the clinical application.

Key words

Immunofluorescence Anti-neutrophil cytoplasmic antibodies Vasculitis Inflammatory bowel disease 



ANCA-associated vasculitis


Anti-nuclear antibodies


Anti-neutrophil cytoplasmic antibodies


Bactericidal permeability increasing protein


Cystic fibrosis


Eosinophilic granulomatosis with polyangiitis


Fluorescein isothiocyanate


Granulomatosis with polyangiitis


Inflammatory bowel disease


Indirect immunofluorescence


Microscopic polyangiitis




Proteinase 3


Primary sclerosing cholangitis


Rheumatoid arthritis


Systemic lupus erythematosus


Small vessel vasculitis



The authors would like to acknowledge Kai Fechner (Euroimmun AG, Germany) for providing the image of formalin-fixed granulocytes.


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • J. J. B. C. van Beers
    • 1
  • J. Vanderlocht
    • 1
  • C. Roozendaal
    • 2
  • J. Damoiseaux
    • 1
  1. 1.Central Diagnostic LaboratoryMaastricht University Medical CentreMaastrichtThe Netherlands
  2. 2.Department of Laboratory Medicine, University Medical Centre GroningenUniversity of GroningenGroningenThe Netherlands

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