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Clinical and Vaccine Immunology, April 2009, p. 464-470, Vol. 16, No. 4
1071-412X/09/$08.00+0 doi:10.1128/CVI.00002-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.
Evaluation of the Combined Application of Ethanol-Fixed and Formaldehyde-Fixed Neutrophil Substrates for Identifying Atypical Perinuclear Antineutrophil Cytoplasmic Antibodies in Inflammatory Bowel Disease
Maria Papp,1*,
Istvan Altorjay,1,
Gabriella Lakos,2
Judit Tumpek,2
Sandor Sipka,2
Tamas Dinya,3
Karoly Palatka,1
Gabor Veres,4
Miklos Udvardy,1 and
Peter Laszlo Lakatos5
2nd Department of Medicine,1 Laboratory of Clinical Immunology,2 1st Department of Surgery, University of Debrecen, Debrecen,3 1st Department of Pediatrics,4 1st Department of Medicine, Semmelweis University, Budapest, Hungary5
Received 3 January 2009/ Returned for modification 21 January 2009/ Accepted 26 January 2009
No clear guidelines for indirect immunofluorescence (IIF) detection and interpretation of antineutrophil cytoplasmic antibodies (ANCA) have been proposed for inflammatory bowel diseases (IBD). We evaluated the reliability of the combined use of ethanol- and formalin-fixed neutrophil substrates to identify atypical perinuclear ANCA (P-ANCA) by IIF under routine laboratory circumstances. A total of 204 IBD patients were assessed with four different fluorescent substrates in two distinct laboratories. Antibodies against myeloperoxidase, proteinase-3, and other specific granule proteins (elastase, lactoferrin, cathepsin G, lysozyme, and bactericidal permeability-increasing protein) were measured by an enzyme-linked immunosorbent assay. The combined application of ethanol- and formalin-fixed slides to detect atypical P-ANCA resulted in a lack of agreement between assays (
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0.39) in the interassay study and moderate agreement in the interobserver study (, 0.42). After atypical and typical P-ANCA patterns were combined, the consensus improved greatly. A total of 26.9% of patients were P-ANCA positive by at least two tests (44.3% of ulcerative colitis [UC] and 13.1% of Crohn's disease [CD] patients; P < 0.0001), while overall ANCA positivity was 22.5% to 34.8%. The combined application of ethanol-fixed and formaldehyde-fixed neutrophil substrates did not facilitate differentiation between P-ANCA and atypical P-ANCA, and the results were not consistent when substrates from different sources were used. Combining all P-ANCA ensures the highest sensitivity and specificity in differentiating UC from CD.
* Corresponding author. Mailing address: 2nd Department of Medicine, University of Debrecen, Nagyerdei krt 98, H-4032 Debrecen, Hungary. Phone and fax: 36 52 31 44 10. E-mail: drpappm{at}yahoo.com
Published ahead of print on 4 February 2009.
M. Papp and I. Altorjay contributed equally to this work, and both should be considered first authors.
Clinical and Vaccine Immunology, April 2009, p. 464-470, Vol. 16, No. 4
1071-412X/09/$08.00+0 doi:10.1128/CVI.00002-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.
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