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

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

Abstract

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 

Abbreviations

AAV

ANCA-associated vasculitis

ANA

Anti-nuclear antibodies

ANCA

Anti-neutrophil cytoplasmic antibodies

BPI

Bactericidal permeability increasing protein

CF

Cystic fibrosis

EGPA

Eosinophilic granulomatosis with polyangiitis

FITC

Fluorescein isothiocyanate

GPA

Granulomatosis with polyangiitis

IBD

Inflammatory bowel disease

IIF

Indirect immunofluorescence

MPA

Microscopic polyangiitis

MPO

Myeloperoxidase

PR3

Proteinase 3

PSC

Primary sclerosing cholangitis

RA

Rheumatoid arthritis

SLE

Systemic lupus erythematosus

SVV

Small vessel vasculitis

Notes

Acknowledgments

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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