Evaluation of the combined application of ethanol-fixed and formaldehyde-fixed neutrophil substrates for identifying atypical perinuclear antineutrophil cytoplasmic antibodies in inflammatory bowel disease

M. Papp, I. Altorjay, Gabriella Lakos, Judit Tumpek, S. Sipka, Tamas Dinya, K. Palatka, G. Verès, M. Udvardy, P. Lakatos

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12 Citations (Scopus)

Abstract

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 (κ ≤0.39) in the interassay study and moderate agreement in the interobserver study (κ, 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.

Original languageEnglish
Pages (from-to)464-470
Number of pages7
JournalClinical and Vaccine Immunology
Volume16
Issue number4
DOIs
Publication statusPublished - ápr. 2009

Fingerprint

Antineutrophil Cytoplasmic Antibodies
Inflammatory Bowel Diseases
Formaldehyde
Neutrophils
Ethanol
Substrates
Indirect Fluorescent Antibody Technique
Ulcerative Colitis
Crohn Disease
Assays
Cathepsin G
Myeloblastin
Immunosorbents
Lactoferrin
Pancreatic Elastase
Muramidase
Peroxidase
Enzyme-Linked Immunosorbent Assay
Guidelines
Sensitivity and Specificity

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Immunology
  • Immunology and Allergy
  • Microbiology (medical)

Cite this

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title = "Evaluation of the combined application of ethanol-fixed and formaldehyde-fixed neutrophil substrates for identifying atypical perinuclear antineutrophil cytoplasmic antibodies in inflammatory bowel disease",
abstract = "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 (κ ≤0.39) in the interassay study and moderate agreement in the interobserver study (κ, 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.",
author = "M. Papp and I. Altorjay and Gabriella Lakos and Judit Tumpek and S. Sipka and Tamas Dinya and K. Palatka and G. Ver{\`e}s and M. Udvardy and P. Lakatos",
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T1 - Evaluation of the combined application of ethanol-fixed and formaldehyde-fixed neutrophil substrates for identifying atypical perinuclear antineutrophil cytoplasmic antibodies in inflammatory bowel disease

AU - Papp, M.

AU - Altorjay, I.

AU - Lakos, Gabriella

AU - Tumpek, Judit

AU - Sipka, S.

AU - Dinya, Tamas

AU - Palatka, K.

AU - Verès, G.

AU - Udvardy, M.

AU - Lakatos, P.

PY - 2009/4

Y1 - 2009/4

N2 - 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 (κ ≤0.39) in the interassay study and moderate agreement in the interobserver study (κ, 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.

AB - 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 (κ ≤0.39) in the interassay study and moderate agreement in the interobserver study (κ, 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.

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