Comparative analysis of different commercial ELISA systems for the detection of anti-neutrophil cytoplasm antibodies in ANCA-associated vasculitides

Clin Exp Rheumatol. 2012 Jan-Feb;30(1 Suppl 70):S66-9. Epub 2012 May 10.

Abstract

Objectives: To assess the diagnostic performance of 11 commercial PR3- and MPO-ANCA ELISA systems (direct, capture and high sensitive [hs] ELISA).

Methods: Sera from 90 patients with AAV (GPA, MPA and CSS) and 20 disease controls (SLE; RA) and healthy individuals were tested for the presence of ANCA by IFT and by different ELISAs for the presence of PR3-and MPO-ANCA, respectively. Furthermore, the binding capacity of the IUIS-CDC reference sera for PR3-/MPO-ANCA in different commercial assays was analysed.

Results: Commercial ELISA kits for PR3-ANCA differed moderately in their sensitivity (from 45% to 62.5%). The highest sensitivity for PR3-ANCA was obtained with hs ELISA (kit A) and capture ELISA (kit N). Testing for MPO-ANCA the highest sensitivity (85%) was obtained with direct ELISA (kit D and I). Specificity was high in all kits. Only three PR3-ANCA commercial kits and three MPO-ANCA kits produced binding at the expected value for the IUIS-CDC reference sera (100 U/ml). In all of the kits, serial dilutions of the reference sera did not yield linearity.

Conclusions: Second (capture) and third (high sensitivity) generation PR3-ANCA ELISA kits are superior to conventional ELISAs. Direct and capture MPO-ANCA ELISAs showed a good overall performance in all kits. Most of the kits have not been standardised to allow their results to be compared.

Publication types

  • Comparative Study
  • Evaluation Study

MeSH terms

  • Anti-Neutrophil Cytoplasmic Antibody-Associated Vasculitis / blood
  • Anti-Neutrophil Cytoplasmic Antibody-Associated Vasculitis / diagnosis*
  • Anti-Neutrophil Cytoplasmic Antibody-Associated Vasculitis / immunology
  • Antibodies, Antineutrophil Cytoplasmic / blood*
  • Biomarkers / blood
  • Case-Control Studies
  • Enzyme-Linked Immunosorbent Assay* / standards
  • Germany
  • Humans
  • Myeloblastin / immunology*
  • Peroxidase / immunology*
  • Predictive Value of Tests
  • Reagent Kits, Diagnostic* / standards
  • Reference Standards
  • Sensitivity and Specificity

Substances

  • Antibodies, Antineutrophil Cytoplasmic
  • Biomarkers
  • Reagent Kits, Diagnostic
  • Peroxidase
  • Myeloblastin