Clinical Rheumatology

, 28:153 | Cite as

A comparative study of IgG second- and third-generation anti-cyclic citrullinated peptide (CCP) ELISAs and their combination with IgA third-generation CCP ELISA for the diagnosis of rheumatoid arthritis

  • Lisiane Maria Enriconi dos Anjos
  • Ivanio Alves Pereira
  • Eleonora d ‘Orsi
  • Andrea Piette Seaman
  • Rufus Watson Burlingame
  • Edelton Flavio Morato
Original Article
First Online:
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Revised:
Accepted:

Abstract

To compare IgG anti-cyclic citrullinated peptide (CCP) enzyme-linked immunosorbent assays (ELISAs) of second (anti-CCP2) and third generations (anti-CCP3) for the diagnosis of rheumatoid arthritis (RA), an IgA CCP3 ELISA was also evaluated. Combinations of the use of the three tests were evaluated. Anti-CCP2 IgG, anti-CCP3 IgG, and anti-CCP3 IgA antibody levels were determined by ELISAs in the serum of 70 patients with rheumatoid arthritis, 34 patients with systemic lupus erythematosus (SLE), and 54 normal subjects. We evaluated the serum levels, diagnostic performance, and the use of a combination of tests for RA diagnosis. Statistical analyses include receiver operating curves (ROCs) and others. The serum levels were higher in RA patients. Sensitivity, specificity, positive predictive value, negative predictive value, positive likelihood ratio (LR), and negative LR were, respectively, 78.6%, 94.3%, 91.7%, 84.7%, 13.8, and 0.23 for anti- CCP2 IgG and 82.9%, 93.2%, 90.6%, 87.2%, 12.2, and 0.18 for anti-CCP3 IgG. These values were better than the same statistical tests for anti-CCP3 IgA. ROC analysis showed that anti-CCP2 IgG and anti-CCP3 IgG had good performance and similar areas. Measuring both IgG and IgA anti-CCP tests increases the specificity if both tests were positive and increases the sensitivity if either test were positive. In our population, anti-CCP2 IgG and anti-CCP3 IgG had good diagnostic performance. Anti-CCP3 IgG had 4.3% more sensitivity than the anti-CCP2 IgG test while sustaining high specificity. This and other studies suggest the development of a dual test—CCP3 IgG and IgA that may be a potential diagnostic tool.

Keywords

Anti-CCP antibodies Clinical laboratory techniques Cyclic citrullinated peptide Rheumatoid arthritis 
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Notes

Acknowledgments

We are grateful to patients and healthy subjects who joined the study for contributed their valuable specimens and to the Hospital Universitario-Universidade Federal de Santa Catarina personnel for the special attention.

Disclosure

Lisiane M. E. Anjos: none; Ivanio A. Pereira: none; Eleonora d‘Orsi: none; and Edelton F. Morato: none. Andrea P. Seaman and Rufus W. Burlingame are employees of Inova Diagnostics, San Diego, CA, USA.

References

  1. 1.
    American College of Rheumatology Ad Hoc Committee on Clinical Guidelines (1996) Guideline arthritis. Arthritis Rheum 39:713–722CrossRefGoogle Scholar
  2. 2.
    American College of Rheumatology Ad Hoc Committee on Clinical Guidelines (2002) Guideline arthritis: 2002 Update. Arthritis Rheum 46:328–346CrossRefGoogle Scholar
  3. 3.
    van Boekel MAM, Vossenaar ER, van den Hoogent FHJ, Venrooij WJ (2002) Autoantibody systems in rheumatoid arthritis: specificity, sensitivity and diagnostic value. Arthritis Res 4:87–93PubMedCrossRefGoogle Scholar
  4. 4.
    Steiner G, Smolen J (2002) Autoantibodies in rheumatoid arthritis and their clinical significance. Arthritis Res 4:S1–S5PubMedCrossRefGoogle Scholar
  5. 5.
    Mageed RA (1996) The RF antigen. In: van Venrooij WJ, Maini RN (eds) Manual of Biological Markers of Disease. Kluwer Academic, Dordrecht, B1.1, pp 1–27Google Scholar
  6. 6.
    Nienhuis RLF, Mandena EA (1964) A new serum factor in patients with rheumatoid arthritis. The antiperinuclear factor. Ann Rheum Dis 23:302–305PubMedCrossRefGoogle Scholar
  7. 7.
    Johnson GD, Carvalho A, Holborow EJ, Goddard DH, Russell G (1981) Antiperinuclear factor and keratin antibodies in rheumatoid arthritis. Ann Rheum Dis 40:263–266PubMedCrossRefGoogle Scholar
  8. 8.
    Hoet RM, Van Venrooij WJ (1992) The antiperinuclear factor and antikeratin antibodies in rheumatoid arthritis. In: Smolen J, Kalden J, Maini RN (eds) Rheumatoid arthritis. Springer, Berlin, pp 299–318Google Scholar
  9. 9.
    Young BJJ, Mallya RK, Leslie RDG, Clark CJM, Hamblin TJ (1979) Anti-keratin antibodies in rheumatoid arthritis. Br Med J 2:97–99PubMedGoogle Scholar
  10. 10.
    Sebbag M, Simon M, Vincent C, Masson-Bessiere C, Girbal E, Duri JJ (1995) The antiperinuclear factor and the so-called antikeratin are the same rheumatoid arthritis-specific autoantibodies. J Clin Invest 95:2672–2679PubMedCrossRefGoogle Scholar
  11. 11.
    Vincent C, Simon M, Sebbag M, Girlbal-Neuhauser E, Durieux JJ, Cantagrel A, Fournie B, Mazieres B, Serre G (1998) Immunoblotting detection of autoantibodies to human epidermis filaggrin: a new diagnostic test for rheumatoid arthritis. J Rheumatol 25:838–846PubMedGoogle Scholar
  12. 12.
    Serre G (2001) Autoantibodies to filaggrin/deiminated fibrin (AFA) are useful for the diagnosis and prognosis of rheumatoid arthritis, and are probably involved in the pathophysiology of the disease. Jt Bone Spine 68:103–105CrossRefGoogle Scholar
  13. 13.
    Schellekens GA, de Jong BAW, van den Hoogen FHJ, van de Putte LBA (1998) Citrulline is an essential constituent of antigenic determinants recognized by rheumatoid arthritis-specific autoantibodies. J Clin Invest 101:273–281PubMedCrossRefGoogle Scholar
  14. 14.
    Girbal-Neuhauser E, Durieux J-J, Arnaud M, Dalbon P, Sebbag M, Vincent C, Simon M, Senshu T, Masson-Bessiere C, Jolivet- Reynaud C, Jolivet M, Serre G (1999) The epitopes targeted by the rheumatoid arthritis-associated antifilaggrin autoantibodies are posttranslationally generated on various sites of (pro)filaggrin by deimination of arginine residues. J Immunol 162:585–594PubMedGoogle Scholar
  15. 15.
    Lapointe E, Dery U, Vaillanciurt F, Menard HA (1999) Rheumatoid sera potentially recognize all citrullinated proteins. Arthritis Rheum 42:86Google Scholar
  16. 16.
    Vittecoq O, Incaurgarat B, Jouen-Beades F, Legoedec J, Letourneur O, Rolland D, Gervasi G, Menard JF, Gayet A, Fardellone P, Daragon A, Jolivet M, Ie Loet X, Tron F (2004) Autoantibodies recognizing citrullinated rat filaggrin in an ELISA using citrullinated and non-citrullinated recombinant proteins as antigens are highly diagnostic for rheumatoid arthritis. Clin Exp Immunol 135:173–180PubMedCrossRefGoogle Scholar
  17. 17.
    Schellekens GA, Visser H, de Jong BAW, Van Den Hoogen FHJ, Hazes JMW, Breedveld FC, Van Venrooij WJ (2000) The diagnostic properties of rheumatoid arthritis antibodies recognizing a cyclic citrullinated peptide. Arthritis Rheum 43:155–163PubMedCrossRefGoogle Scholar
  18. 18.
    Van Gaalen FA, Visser H, Huizinga TW (2005) A comparison of the diagnostic accuracy and prognostic value of the first and second anti-cyclic citrullinated peptides (CCP1 and CCP2) autoantibody tests for rheumatoid arthritis. Ann Rheum Dis 64:1510–1512PubMedCrossRefGoogle Scholar
  19. 19.
    Saraux A, Berthelot JM, Devauchelle V, Bendaoud B, Chalès G, Le Henaff C, Thorel JB, Hoang S, Jousse S, Baron D, Le Goff P, Youinou P (2004) Value of antibodies to citrulline-containing peptides for diagnosing early rheumatoid arthritis. J Rheumatol 30:2535–2539Google Scholar
  20. 20.
    Sauerland U, Becker H, Seide M, Schotte H, Willeke P, Schorat A, Schluter B, Domschke W, Gaubitz M (2005) Clinical utility of the anti-CCP assay: experiences with 700 patients. Ann NY Acad Sci 1050:314–318PubMedCrossRefGoogle Scholar
  21. 21.
    Vallbracht I, Rieber J, Oppermann M, Forger F, Siebert U, Helmke K (2004) Diagnostic and clinical value of anti-cyclic citrulinated peptide antibodies compared with rheumatoid factor isotypes in rheumatoid arthritis. Ann Rheum Dis 63:1079–1084PubMedCrossRefGoogle Scholar
  22. 22.
    Dubrous P, Gardet V, Hugard L (2005) Value of anti-cyclic citrullinated peptides antibodies in comparison with rheumatoid factor for rheumatoid arthritis diagnosis. Pathol Biol 53:63–67PubMedCrossRefGoogle Scholar
  23. 23.
    Silveira IG, Burlingame RW, von Mühlen CA, Bender AL, Staub HL (2007) Anti-CCP antibodies have more diagnostic impact than rheumatoid factor (RF) in a population tested for RF. Clin Rheumatol 26:1883–1889PubMedCrossRefGoogle Scholar
  24. 24.
    Vallbracht I, Helmke K (2005) Additional diagnostic and clinical value of anti-cyclic citrullinated peptide antibodies compared with rheumatoid factor isotypes in rheumatoid arthritis. Autoimmun Rev 4:389–394PubMedCrossRefGoogle Scholar
  25. 25.
    Solanki K, Spellerberg M, Chapman P, Moller P, O’Donnell J (2004) Anti-cyclic citrullinated antibodies: complementary to IgM rheumatoid factor in the early diagnosis of rheumatoid arthritis. N Z Med J 117:1097Google Scholar
  26. 26.
    Araki C, Hayashi N, Moriyama M, Morinobu S, Mukai M, Koshiba M, Kawano S, Kumagai S (2004) Usefulness of anti-cyclic citrullinated peptide antibodies (anti-CCP) for the diagnosis of rheumatoid arthritis. Rinsho Byori 52:966–972PubMedGoogle Scholar
  27. 27.
    Avouac J, Gossec L, Dougados M (2006) Diagnostic and predictive value of anti-cyclic citrullinated protein antibodies in rheumatoid arthritis: a systematic literature review. Ann Rheum Dis 65:845–851PubMedCrossRefGoogle Scholar
  28. 28.
    Nishimura K, Sugiyama D, Kogata Y, Tsuji G, Nakazawa T, Kawano S, Saigo K, Morinobu A, Koshiba M, Kuntz KM, Kamae I, Kumagai S (2007) Meta-analysis: diagnostic accuracy of anti-cyclic citrullinated peptide antibody and rheumatoid factor for rheumatoid arthritis. Ann Intern Med 146:797–808PubMedGoogle Scholar
  29. 29.
    Girelli F, Foscbi FG, Bedescbi E, Calderoni V, Stefanini GF, Martinelli MG (2004) Is Anti Cyclic citrullinated peptide a useful laboratory test for the diagnosis of rheumatoid arthritis? Allerg Immunol 36:127–130Google Scholar
  30. 30.
    Berglin E, Padyukov L, Sundin U, Hallmans G, Stenlund H, van Venrooij WJ, Klareskog L, Dahlqvist SR (2004) A combination of autoantibodies to cyclic citrullinated peptide (CCP) and HLA DRB1 locus antigens is strongly associated with future onset of rheumatoid arthritis. Arthritis Res Ther 6:R303–R308PubMedCrossRefGoogle Scholar
  31. 31.
    Nielen MJ, van Shaardenburg D, Reesink HW, van de Stadt RJ, van der Horst-Bruinima IE, Dekoning MHMT, Habibuw MR, Vandenbroucke JP, Dijkmans BAC (2004) Specific autoantibodies precede the symptoms of Rheumatoid Arthritis. Arthritis Rheum 50:380–386PubMedCrossRefGoogle Scholar
  32. 32.
    Van Gaalen FA, Linn-Rasker SP, van Venrooij WJ, de Jong BA, Breedveld FC, Verweij CC, Toes REM, Huizinga TWJ (2004) Autoantibodies to cyclic citrullinated peptides predict progression to rheumatoid arthritis in patients with undifferentiated arthritis: A prospective cohort study. Arthritis Rheum 50:709–715PubMedCrossRefGoogle Scholar
  33. 33.
    Rormelid J, Wick MC, Lampa J, Lindblad S, Nordmark B, Klareskog L, van Vollenhoven RF (2005) Longitudinal analysis of citrullinated protein/peptide antibodies (anti-CCP) during 5 year follow up in early rheumatoid arthritis: anti-CP status predicts worse disease activity and greater radiological progression. Ann Rheum Dis 64:1744–1749CrossRefGoogle Scholar
  34. 34.
    De Rycke L, Peene I, De Keyser F (2004) Rheumatoid factor and anticitrullinated protein antibodies in rheumatoid arthritis: diagnostic value, associations with radiological progression rate, and extra-articular manifestations. Ann Rheum Dis 63:1587–1593PubMedCrossRefGoogle Scholar
  35. 35.
    van Jaarsveld CD, ter Borg EJ, Jacobs JW, Schellekens GA, Gmelig-Meyling FH, van Booma-Frankfort C, de Jong BA, van Venrooij WJ, Bijlsma JW (1999) The prognostic value of the antiperinuclear factor, anticitrullinated peptide antibodies and rheumatoid factor in early rheumatoid arthritis. Clin Exp Rheumatol 17:689–697PubMedGoogle Scholar
  36. 36.
    Verpoort KN, Jol-van der Zijde CM, Papendrecht-van der Voort EA, Ioan-Facsinay A, Drijfhout JW, van Tol MJ, Breedveld FC, Huizinga TW, Toes RE (2006) Isotype distribution of anti-cyclic citrullinated peptide antibodies in undifferentiated arthritis and rheumatoid arthritis reflects an ongoing immune response. Arthritis Rheum 54:3799–3808PubMedCrossRefGoogle Scholar
  37. 37.
    Santiago M, Baron M, Miyachi K, Fritzler MJ, Abu-Hakima M, Leclercq S, Bell M, Hudson M, Mathieu JP, Taillefer S, Jones N, Docherty P, Khraishi M, Markland J, Pope J, Robinson D, Smith D, Sutton E (2008) A comparison of the frequency of antibodies to cyclic citrullinated peptides using a third generation anti-CCP assay (CCP3) in systemic sclerosis, primary biliary cirrhosis and rheumatoid arthritis. Clin Rheumatol 27:77–83PubMedCrossRefGoogle Scholar
  38. 38.
    Wu R, Shovman O, Zhang Y, Gilburd B, Zandman-Goddard G, Shoenfeld Y (2007) Increased prevalence of anti-third generation cyclic citrullinated peptide antibodies in patients with rheumatoid arthritis and CREST syndrome. Clin Rev Allergy Immunol 32:47–56PubMedCrossRefGoogle Scholar
  39. 39.
    American College of Rheumatology (1997) Revised Criteria for the Classification of Systemic Lupus Erythematosus. Arthritis Rheum 40:1725Google Scholar
  40. 40.
    Classification Criteria of Rheumatoid Arthrites, 1987. www.rheumatology.org/publications/classification/ra/1987 (accessed Dec 2007)
  41. 41.
    Martinez JB, Valero JS, Bautista AJ, Restrepo JF, Matteson EL, Rondon F, Iglesias-Gamarra A (2007) Erosive arthropathy: clinical variance in lupus erythematosus and association with anti-CCP case series and review of the literature. Clin Exp Rheumatol 25:47–53PubMedGoogle Scholar

Copyright information

© Clinical Rheumatology 2008

Authors and Affiliations

  • Lisiane Maria Enriconi dos Anjos
    • 1
  • Ivanio Alves Pereira
    • 2
  • Eleonora d ‘Orsi
    • 2
  • Andrea Piette Seaman
    • 3
  • Rufus Watson Burlingame
    • 3
  • Edelton Flavio Morato
    • 2
  1. 1.Universidade do Sul de Santa Catarina (UNISUL)TubaraoBrazil
  2. 2.Universidade Federal de Santa Catarina (UFSC)FlorianopolisBrazil
  3. 3.Inova DiagnosticsSan DiegoUSA

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