Simultaneous automated screening and confirmatory testing for vasculitis-specific ANCA

Mandy Sowa, Kai Grossmann, Ilka Knütter, Rico Hiemann, Nadja Röber, Ursula Anderer, Elena Csernok, Dimitrios P. Bogdanos, Maria Orietta Borghi, Pier Luigi Meroni, Peter Schierack, Dirk Reinhold, Karsten Conrad, Dirk Roggenbuck

Research output: Contribution to journalArticlepeer-review


Anti-neutrophil cytoplasmic antibodies (ANCA) are the serological hallmark of small vessel vasculitis, so called ANCA associated vasculitis. The international consensus requires testing by indirect immunofluorescence (IIF) on human ethanolfixed neutrophils (ethN) as screening followed by confirmation with enzyme-linked immunosorbent assays (ELISAs). This study evaluates the combination of cell- and microbead-based digital IIF analysis of ANCA in one reaction environment by the novel multiplexing CytoBead technology for simultaneous screening and confirmatory ANCA testing. Sera of 592 individuals including 118 patients with ANCA-associated vasculitis, 133 with rheumatoid arthritis, 49 with infectious diseases, 77 with inflammatory bowel syndrome, 20 with autoimmune liver diseases, 70 with primary sclerosing cholangitis and 125 blood donors were tested for cytoplasmic ANCA (C-ANCA) and perinuclear ANCA (P-ANCA) by classical IIF and ANCA to proteinase 3 (PR3) and myeloperoxidase (MPO) by ELISA. These findings were compared to respective ANCA results determined by automated multiplex CytoBead technology using ethN and antigen-coated microbeads for microbead immunoassays. There was a good agreement for PR3-and MPO-ANCA and a very good one for P-ANCA and C-ANCA by classical and multiplex analysis (Cohen's kappa [κ] = 0.775, 0.720, 0.876, 0.820, respectively). The differences between classical testing and CytoBead analysis were not significant for PR3-ANCA, P-ANCA, and C-ANCA (p

Original languageEnglish
Article numbere107743
JournalPLoS One
Issue number9
Publication statusPublished - Sept 16 2014

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)


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