Jump to ContentJump to Main Navigation
Show Summary Details
In This Section

Clinical Chemistry and Laboratory Medicine (CCLM)

Published in Association with the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)

Editor-in-Chief: Plebani, Mario

Ed. by Gillery, Philippe / Lackner, Karl J. / Lippi, Giuseppe / Melichar, Bohuslav / Payne, Deborah A. / Schlattmann, Peter / Tate, Jillian R.

12 Issues per year


IMPACT FACTOR 2016: 3.432

CiteScore 2016: 2.21

SCImago Journal Rank (SJR) 2015: 0.873
Source Normalized Impact per Paper (SNIP) 2015: 0.982

Online
ISSN
1437-4331
See all formats and pricing
In This Section
Volume 42, Issue 10 (Oct 2004)

Issues

Analytical and diagnostic accuracy of the EliA™ automated enzyme fluoroimmunoassay for antineutrophil cytoplasmic autoantibody detection

Danilo Villalta
  • Immunologia Clinica e Virologia, DML, A.O. “S. Maria degli Angeli”, Pordenone, Italy
/ Elio Tonutti
  • Dipartimento di Diagnostica di Laboratorio, A.O. “S. Maria della Misericordia”, Udine, Italy
/ Marilina Tampoia
  • Laboratorio di Patologia Clinica, Policlinico, Bari, Italy
/ Nicola Bizzaro
  • Laboratorio di Patologia Clinica, Ospedale Civile, San Donà di Piave, Italy
/ Wolfgang Papisch
  • Pharmacia Diagnostics, Freiburg, Germany
/ Renato Tozzoli
  • Dipartimento di Medicina di Laboratorio, Ospedale Civile, Latisana, Italy
/ Sergio Stella
  • Immunologia Clinica e Virologia, DML, A.O. “S. Maria degli Angeli”, Pordenone, Italy
Published Online: 2011-09-21 | DOI: https://doi.org/10.1515/CCLM.2004.236

Abstract

Anti-proteinase 3 antineutrophil cytoplasmic antibodies (PR3-ANCA) and anti-myeloperoxidase antibodies (MPO-ANCA) are considered important serological markers for several forms of idiopathic systemic vasculitis. The aim of the study was to verify the analytical and clinical performance of a new automated enzyme fluoroimmunoassay, the EliA system, for PR3-ANCA and MPO-ANCA detection.

For this purpose the sera of 52 consecutive well-defined patients with a clinical diagnosis of Wegener’s granulomatosis (WG) (n = 29) or microscopic polyangiitis (MPA) (n = 23), and 70 controls suffering from connective tissue disease (25 systemic lupus erythematosus, 25 Sjögren’s syndrome and 20 rheumatoid arthritis) were tested for PR3-ANCA and MPO-ANCA with the EliA assay (Pharmacia Diagnostics, Freiburg, Germany). For comparison purposes, the same sera were also tested by indirect immunofluorescence, another direct immunometric assay (Varelisa, Pharmacia Diagnostics) and a capture PR3-ANCA (Wieslab AB, Lund, Sweden) method.

Both the EliA PR3-ANCA and MPO-ANCA assays showed between- and within-assay precision of < 10%. The dilution test gave straight lines (r2 = 0.998) for both antibody assays. The recovery ranged from 97.9% to 102.7% for PR3-ANCA and from 84.9% to 91.4% for MPO-ANCA.

There was a high positive correlation between the EliA and Varelisa methods for quantitative detection of MPO-ANCA levels (r2 = 0.949) and a lower correlation for PR3-ANCA (r2 = 0.771). Conversely, poor correlation was observed between EliA PR3-ANCA and capture PR3-ANCA (r2 = 0.537). The overall sensitivity and specificity of EliA PR3-ANCA and MPO-ANCA for the vasculitides considered in this study were 82.7% and 97.2%, respectively, with a positive predictive value of 96.6% and a negative predictive value of 84.9%. Comparison of the results obtained with the indirect immunofluorescence, Varelisa and capture PR3-ANCA methods showed that the indirect immunofluorescence assay is the most sensitive method for the diagnosis of vasculitis (88.5%), but the least specific (94.3%); the EliA method is slightly more specific (97.2%) than the Varelisa method (95.7%), and also slightly more sensitive (82.7% vs. 80.8%). Capture PR3-ANCA proved to be the most sensitive method for detection of anti-proteinase 3 antibodies in WG (89.7% vs. 86.2% EliA and 79.3% Varelisa).

In conclusion, the EliA MPO-ANCA and PR3-ANCA methods provide good diagnostic accuracy and excellent analytical accuracy, which, in association with the practicality of the automated EliA system, make this method a useful tool for the diagnosis of ANCA-associated vasculitides.

Keywords: anti-proteinase 3 antibodies; anti-myeloperoxidase antibodies; Wegener’s granulomatosis; microscopic polyangiitis; enzyme fluoroimmunoassay

References

  • 1

    Jennette JC, Falk RJ. Antineutrophil cytoplasmic autoantibodies and associated diseases: a review. Am J Kidney Dis 1990; 15:517–29. [Crossref]

  • 2

    Hoffman GS, Specks U. Antineutrophil cytoplasmic autoantibodies. Arthritis Rheum 1998; 41:1521–37. [Crossref]

  • 3

    Hagen EC, Ballieux BEPB, van Es LA, Daha MR, van der Woude FJ. Anti-neutrophil cytoplasmic autoantibodies (ANCA). A review of the antigens involved, the assays, the clinical and possible pathogenetic consequences. Blood 1993; 81:1996–2002.

  • 4

    Wieslander J. How are antineutrophil cytoplasmic antibodies detected? Am J Kidney Dis 1991; 18:154–8. [Crossref]

  • 5

    Ludemann J, Utech B, Gross WL. Anti-cytoplasmic antibodies in Wegener’s granulomatosis are directed against proteinase 3. Adv Exp Med Biol 1991; 297:141–50.

  • 6

    Falk RJ, Jennette JC. Anti neutrophil cytoplasmic autoantibodies with specificity for myeloperoxidase in patients with systemic vasculitis and idiopathic necrotizing and crescentic glomerulonephritis. N Engl J Med 1988; 318:1651–7. [Crossref]

  • 7

    Coremans IEM, Hagen EC, Daha MR, van der Woude FJ, van der Voort EAM, Kleijburgvanderkeur C, et al. Anti-lactoferrin antibodies in patients with rheumatoid arthritis are associated with vasculitis. Arthritis Rheum 1992; 32:1466–75. [Crossref]

  • 8

    Cohen Tervaert JW, Mulder L, Stegeman C, Elema J, Huitema M, The H, et al. Occurrence of autoantibodies to human leucocyte elastase in Wegener’s granulomatosis and other inflammatory disorders. Ann Rheum Dis 1993; 52:115–20. [Crossref]

  • 9

    Zhao Mh, Jones SJ, Lockwood CM. Bactericidal/permeability-increasing protein (BPI) is an important antigen for anti-neutrophil cytoplasmic autoantibodies (ANCA) in vasculitis. Clin Exp Immunol 1995; 99:49–56. [Crossref]

  • 10

    Halbwachs Mecarelli L, Nusbaum P, Noel LH, Reumax D, Erlinger S, Grunfeld JP, et al. Antineutrophil cytoplasmic antibodies (ANCA) directed against cathepsin G in ulcerative colitis, Crohn’s disease and primary sclerosing cholangitis. Clin Exp Immunol 1992; 90:79–84. [Crossref]

  • 11

    Wiik A, Jensen E, Friis J. Granulocyte-specific antinuclear factors in synovial fluids and sera from patients with rheumatoid arthritis. Ann Rheum Dis 1974; 33:515–22. [Crossref]

  • 12

    Hagen EC, Daha MR, Hermans J, Andrassay K, Csernok E, Gaskin G, et al., for the EC/BCR Project for ANCA Assay Standardization. Diagnostic value of standardized assays for anti-neutrophil cytoplasmic antibodies in idiopathic systemic vasculitis. Kidney Int 1998; 53:743–53. [Crossref]

  • 13

    Sinico RA, Radice A, Pozzi C, Ferrario F, Ardigo G and the Italian Group of Renal Immunopathology. Diagnostic significance and antigen specificity of antineutrophil cytoplasmic antibodies in renal diseases. Nephrol Dial Transplant 1994; 9:505–10.

  • 14

    Savige J, Gillis D, Benson E, Davies D, Esnault V, Falk RJ, et al. International Consensus Statement on Testing and Reporting of antineutrophil cytoplasmic antibodies (ANCA). Am J Clin Pathol 1999; 111:507–13. [Crossref]

  • 15

    Jennette JC, Falk RJ, Andrassay K, Bacon P, Churg J, Gross WL, et al. Nomenclature of systemic vasculitides: Proposal of an international consensus conference. Arthritis Rheum 1994; 37:187–92. [Crossref]

  • 16

    Leavitt RY, Fauci AS, Bloch DA, Michel BA, Hunder CG, Arend WP, et al. The American College of Rheumatology 1990 criteria for the classification of Wegener’s granulomatosis. Arthritis Rheum 1990; 33:1101–7. [Crossref]

  • 17

    Tan EM, Cohen AS, Fries JF, Masi AT, McShane DJ, Rothfield NF, et al. The 1982 revised criteria for the classification of systemic lupus erythematosus. Arthritis Rheum 1982; 25:1271–7. [Crossref]

  • 18

    Vitali C, Bombardieri S, Moutsopoulos HM, Balestrieri G, Bencivelli W, Bernstein RM, et al. Assessment of the European classification criteria for Sjögren’s syndrome in a series of clinically defined cases: results of a prospective multicentre study. The European Study Group in Diagnostic Criteria for Sjögren’s Syndrome. Ann Rheum Dis 1996; 55:116–21. [Crossref]

  • 19

    Arnett FC, Edworthy SM, Bloch DA, McShane DJ, Fries JF, Cooper NS, et al. The American Rheumatism Association 1987 revised criteria for the classification of rheumatoid arthritis. Arthritis Rheum 1988; 31:315–24. [Crossref]

  • 20

    Wiik A, Rasmussen N, Wieslander J. Methods to detect autoantibodies to neutrophilic granulocytes. In: van Venrooij WJ, Maini RN, editors. Manual of biological markers of disease, vol A 9. Dordrecht: Kluwer Academic Publishers, 1993:1–14.

  • 21

    Nölle B, Specks U, Lüdemann J, Rohrbach M, Deremee R, Gross W. Anticytoplasmic autoantibodies: their immunodiagnostic value in Wegener granulomatosis. Ann Int Med 1989; 111:28–40. [Crossref]

  • 22

    Falk RJ, Jennette JC. ANCA small-vessel vasculitis. J Am Soc Nephrol 1997; 8:314–22. [Web of Science]

  • 23

    Hoffman GS, Kerr GS, Leavitt RY, Hallahan CW, Lebovics RS, Travis WD, et al. Wegener granulomatosis: an analysis of 158 patients. Ann Int Med 1992; 116:488–98. [Crossref]

  • 24

    Westman KWA, Selga D, Bygren P, Segelmark M, Baslund B, Wiik A, et al. Clinical evaluation of a capture ELISA for detection of proteinase 3 antineutrophil cytoplasmic antibody. Kidney Int 1998; 53:1230–6. [Crossref]

  • 25

    Wang G, Csernok E, de Groot K, Gross WL. Comparison of eight commercial kits for quantitation of antineutrophil cytoplasmic antibodies (ANCA). J Immunol Methods 1997; 208:203–11. [Crossref]

  • 26

    Pollock W, Dunster K, Roland JM, Koh H, Savige J. A comparison of commercial and in-house ELISAs for antineutrophil cytoplasmic antibodies directed against proteinase 3 and myeloperoxidase. Pathology 1999; 31:38–43. [Crossref]

  • 27

    Csernok E, Ahluist D, Ullrich S, Gross WL. A critical evaluation of commercial immunoassays for antineutrophil cytoplasmic antibodies directed against proteinase 3 and myeloperoxidase in Wegener’s granulomatosis and microscopic polyangiitis. Rheumatology 2002; 41:1313–7. [Crossref]

  • 28

    Lim LCL, Taylor J, Schmitz JL, Folds JD, Wilkman AS, Falk RJ, et al. Diagnostic usefulness of antineutrophil cytoplasmic autoantibody serology. Comparative evaluation of commercial indirect fluorescent antibody kits and enzyme immunoassay kits. Am J Clin Pathol 1999; 111:363–9. [Crossref]

  • 29

    Jethwa HS, Nachman PH, Falk RJ, Jennette JC. False-positive myeloperoxidase binding activity due to DNA/anti-DNA antibody complexes: a source for analytical error in serologic evaluation of anti-neutrophil cytoplasmic autoantibodies. Clin Exp Immunol 2000; 121:544–50. [Crossref]

  • 30

    Van der Woude FJ, Rasmussen N, Lobatto S, Wiik A, Permin H, van Es LA, et al. Autoantibodies against neutrophils and monocytes: tool for diagnosis and marker of disease activity in Wegener’s granulomatosis. Lancet 1985; i:425–9.

  • 31

    Kerr GS, Fleisher TA, Hallahan CW, Leavitt RY, Fauci AS, Hoffman GS. Limited prognostic value of changes in antineutrophil cytoplasmic antibody titer in patients with Wegener’s granulomatosis. Arthritis Rheum 1993; 36:365–71. [Crossref]

  • 32

    Davenport A, Lock RJ, Wallington T. Clinical significance of serial measurement of autoantibodies to neutrophil cytoplasm using a standard indirect immunofluorescence test. Am J Nephrol 1995; 15:201–7. [Crossref]

  • 33

    Specks U, Wheatley CL, McDonald TJ, Rohrbach MS, De Remee RA. Anticytoplasmic autoantibodies in the diagnosis and follow up of Wegener’s granulomatosis. Mayo Clin Proc 1989; 64:28–36. [Crossref]

  • 34

    Tervaert JW, van der Woude FJ, Fauci AS, Ambrus JL, Velosa J, Keane WF, et al. Association between active Wegener’s granulomatosis and anticytoplasmic antibodies. Arch Intern Med 1989; 149:2461–5. [Crossref]

  • 35

    Tervaert JW, Huitema MG, Hene RJ, Sluiter WJ, The TH, van der Hem GK, et al. Prevention of relapses in Wegener’s granulomatosis by treatment based on antineutrophil cytoplasmic antibody titre. Lancet 1990; 336:709–11. [Crossref]

  • 36

    Egner W, Chapel HR. Titration of antibodies against neutrophil cytoplasmic antigens is useful in monitoring disease activity in systemic vasculitides. Clin Exp Immunol 1990; 82:244–9. [Crossref]

  • 37

    Boomsma MM, Stegeman CA, van der Leij, Oost W, Hermans J, Kallenberg CG, et al. Prediction of relapses in Wegener’s granulomatosis by measurement of antineutrophil cytoplasmic antibody levels: a prospective study. Arthritis Rheum 2000; 43:2025–33. [Crossref]

  • 38

    Han Wk, Choi HK, Roth RM, McCluskey RT, Niles JC. Serial ANCA titers: useful tool for prevention of relapses in ANCA-associated vasculitis. Kidney Int 2003; 63:1079–85. [Crossref]

About the article

Received: 2004-06-04

Accepted: 2004-08-20

Published Online: 2011-09-21

Published in Print: 2004-10-01



Citation Information: Clinical Chemistry and Laboratory Medicine (CCLM), ISSN (Online) 1437-4331, ISSN (Print) 1434-6621, DOI: https://doi.org/10.1515/CCLM.2004.236. Export Citation

Citing Articles

Here you can find all Crossref-listed publications in which this article is cited. If you would like to receive automatic email messages as soon as this article is cited in other publications, simply activate the “Citation Alert” on the top of this page.

[2]
Renato Tozzoli
Autoimmunity Reviews, 2007, Volume 6, Number 6, Page 334

Comments (0)

Please log in or register to comment.
Log in