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A novel ELISA using a mixture of human native and recombinant proteinase-3 significantly improves the diagnostic potential for ANCA-associated vasculitis
  1. Jan Damoiseaux (jdam{at}limm.azm.nl)
  1. University Hospital Maastricht, Netherlands
    1. Cornelia Dähnrich (c.dahnrich{at}euroimmun.de)
    1. EUROIMMUN AG, Lübeck, Germany
      1. Anke Rosemann (a.rosemann{at}euroimmun.de)
      1. EUROIMMUN AG, Lübeck, Germany
        1. Christian Probst (c.probst{at}euroimmun.de)
        1. EUROIMMUN AG, Lübeck, Germany
          1. Lars Komorowski (l.komorowski{at}euroimmun.de)
          1. EUROIMMUN AG, Lübeck, Germany
            1. Coen A Stegeman (c.a.stegeman{at}int.umcg.nl)
            1. Department of Nephrology, UMCG, Groningen, Netherlands
              1. Karl Egerer (karl.egerer{at}charite.de)
              1. Department for Rheumatology and Clinical Immunology, Charite University of Medicine, Berlin, Germany
                1. Falk Hiepe (falk.hiepe{at}charite.de)
                1. Department for Rheumatology and Clinical Immunology, Charite University of Medicine, Berlin, Germany
                  1. Pieter Van Paassen (p.vanpaassen{at}immuno.unimaas.nl)
                  1. University Hospital Maastricht, Netherlands
                    1. Winfried Stöcker (w.stocker{at}euroimmun.de)
                    1. EUROIMMUN AG, Luebeck, Germany
                      1. Wolfgang Schlumberger (w.schlumberger{at}euroimmun.de)
                      1. EUROIMMUN AG, Luebeck, Germany
                        1. Jan Willem Cohen Tervaert (jw.cohentervaert{at}immuno.unimaas.nl)
                        1. University Hospital Maastricht, Netherlands

                          Abstract

                          Anti-neutrophil cytoplasmic antibodies (ANCA) with a C-ANCA or P-ANCA pattern are detected in ANCA-associated vasculitis (AAV). While in most AAV patients a C-ANCA pattern is due to reactivity with proteinase-3 (PR3)-ANCA, some C-ANCA positive sera do not react with PR3.

                          Objective: The development and evaluation of a direct ELISA for PR3-ANCA with increased sensitivity.

                          Methods: A mixture of human native (hn) and human recombinant (hr) PR3 was used as antigen coating. The resulting ELISA (Anti-PR3-hn-hr) was compared with ELISAs using directly coated hn-PR3 or hr-PR3, as well as with a hn-PR3 capture ELISA. Assay characteristics were determined in AAV patients (n=248), with special attention for those patients with C-ANCA (n=132), as well as disease controls (n=585) and healthy controls (n=429). Additionally, for prediction of relapses serial samples of 46 PR3-AAV patients were analysed.

                          Results: At a predefined specificity of 99% both ELISAs containing hr-PR3 revealed a substantial increase from 14.4 to 18.2% in sensitivity. For prediction of relapses by rises in PR3-ANCA titers the capture ELISA was most optimal (odds ratio 12.5). With an odds ratio of 8.9 the novel Anti-PR3-hn-hr ELISA was second best.

                          Conclusion: Due to the very high sensitivity of the novel Anti-PR3-hn-hr ELISA for detection of PR3-ANCA in C-ANCA positive samples of AAV patients this assay has an excellent diagnostic performance. This feature is combined with a good predictability of clinical relapses in PR3-AAV patients. These characteristics challenge the dogma that for detection of PR3-ANCA capture ELISAs are superior for diagnosis and follow-up.

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