Skip to main content
Journal of Clinical Laboratory Analysis logoLink to Journal of Clinical Laboratory Analysis
. 2010 Jul 12;24(4):230–236. doi: 10.1002/jcla.20391

Determination of cut‐off titers and agreement between immunofluorescence and immunoblotting methods for detecting antinuclear antibodies in children

G Aksu 1,, N Gulez 1, E Azarsız 1, N Karaca 1, N Kutukçuler 1
PMCID: PMC6647563  PMID: 20626019

Abstract

Detection of antinuclear antibodies (ANA) is a diagnostic adjunct in patients with suspected autoimmune connective tissue diseases, and various detection methods are in use. The aim of this study was to analyze the agreement between the ANA immunoflourescence (IF) and immunoblotting (IB) methods and determine cut‐off for children subjects in a laboratory setting. We evaluated 729 serum samples that were analyzed by both ANA IF and IB. The results were evaluated by χ2 test and, for agreement, κ index was used. Frequencies determined for both 1:40–1:100 cut‐off titers of ANA IF in relation to IB testing supported the idea that 1:100 starting dilution should be recommended in children subjects for ANA IF method and antigen specific immunoblot testing was needed, especially for some of the ANA IF negative samples. Agreement between the two methods, especially with homogenous, granular, and nucleolar ANA IF patterns, was statistically significant. J. Clin. Lab. Anal. 24:230–236, 2010. © 2010 Wiley‐Liss, Inc.

Keywords: ANA, immunoflourescence, immunoblotting, cut‐off titer, children

REFERENCES

  • 1. Tan EM, Feltkamp TEW, Smolen JS. Range of antinuclear antibodies in “healthy” individiuals. Arthritis Rheum 1997;40:1601. [DOI] [PubMed] [Google Scholar]
  • 2. Kumar Y, Bhatia A, Minz RW. Antinuclear antibodies and their detection methods in diagnosis of connective tissue diseases: A journey revisited. Diagn Pathol 2009;4:1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 3. Muro Y. Antinuclear antibodies. Autoimmunity 2005;38:3–9. [DOI] [PubMed] [Google Scholar]
  • 4. Kavanaugh A, Tomar R, Reveille J, Solomon DH, Homburger HA. Guidelines for clinical use of the antinuclear antibody test and tests for specific autoantibodies to nuclear antigens. Arch Pathol Lab Med 2000;124:71–81. [DOI] [PubMed] [Google Scholar]
  • 5. Hayashi N, Kawamoto T, Mukai M, et al. Detection of antinuclear antibodies by use of an enzyme immunoassay with nuclear Hep 2 cell extract and recombinant antigens: Comparison with immunofluorescence assay in 307 patients. Clin Chem 2001;47:1649–1659. [PubMed] [Google Scholar]
  • 6. Gonzalez C, Martin T, Arroyo, T, Isıdoro MG, Navajo J, Buitrago JMG. Comparison and variation of different methodologies for the detection of autoantibodies to nuclear antigens (ANA). J Clin Lab Anal 1997;11:388–392. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 7. Griesmacher A, Peichl. Autoantibodies associated with rheumatic diseases. Clin Chem Lab Med 2001;39:189–208. [DOI] [PubMed] [Google Scholar]
  • 8. Eissfeller P, Sticherling M, Scholz D, et al. Comparison of different test systems for simultaneous autoantibody detection in connective tissue diseases. Ann N Y Acad Sci 2005;1050:327–339. [DOI] [PubMed] [Google Scholar]
  • 9. Thomas C, Robinson JA. The antinuclear antibody test. When is a positive result clinically relevant? Postgrad Med 1993;94:55–66. [PubMed] [Google Scholar]
  • 10. Peene I, Meheus L, Veys EM, De Keyser F. Detection and identification of antinuclear antibodies (ANA) in a large and consecutive cohort of serum samples referred for ANA testing. Ann Rheum Dis 2001;60:1131–1136. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 11. Bonaguri C, Melegari A, Dall'aglio P, et al. An Italian multicenter study for application of a diagnostic algorithm in autoantibody testing. Ann N Y Acad Sci 2009;1173:124–129. [DOI] [PubMed] [Google Scholar]
  • 12. Ghosh P, Duvivedi S, Naik S, et al. Antinuclear antibodies by indirect immunofluorescence: Optimum screening dilution for diagnosis of systemic lupus erythematosus. Indian J Med Res 2007;126:34–38. [PubMed] [Google Scholar]
  • 13. Sack U, Conrad K, Csernok E, et al. Autoantibody detection using indirect immunofluorescence on hep 2 cells. Ann N Y Acad Sci 2009;1173:166–173. [DOI] [PubMed] [Google Scholar]
  • 14. Mcghee JL, Kickingbird LM, Jarvis JN. Clinical utility of antinuclear antibody tests in children. BMC Pediatr 2004;4:13. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 15. Hilario MOE, Len CA, Roja SC, Terreri MT, Almeida G, Andrade LEC. Frequency of antinuclear antibodies in healthy children and adolescents. Clin Pediatr (Phila) 2004;43:637. [DOI] [PubMed] [Google Scholar]
  • 16. Egner W. The use of laboratory tests in the diagnosis of SLE. J Clin Pathol 2000;53:424. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 17. Kern P, Kron M, Hiesche K. Measurement of antinuclear antibodies: Assessment of different test systems. Clin Diagn Lab Immunol 2000;7:72–78. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 18. Bagnasco M, Grassia L, Pesce G. The management of the patient with unexpected autoantibody positivity. Autoimmun Rev 2007;6:347–353. [DOI] [PubMed] [Google Scholar]
  • 19. Bizzaro N, Tozzoli R, Tanutti E, et al. Variability between methods to determine ANA, anti‐dsDNA and anti‐ENA antibodies; a collaborative study with the biomedical industry. J Immunol Methods 1998;219:99–107. [DOI] [PubMed] [Google Scholar]
  • 20. Hoffman IEA, Peene I, Veys EM, Keyser FD. Detection of specific antinuclear reactivities in patients with negative anti‐nuclear antibody immunofluorescence screening tests. Clin Chem 2002;48:2171–2176. [PubMed] [Google Scholar]
  • 21. Kang I, Siperstein R, Quan T, Breitenstein ML. Utility of age, gender, ANA titer and pattern as predictors of anti‐ENA and‐dsDNA antibodies. Clin Rheumatol 2004;23:509–515. [DOI] [PubMed] [Google Scholar]
  • 22. Perilloux BC, Shetty AK, Leiva LE, Gedalia A. Antinuclear antibody (ANA) and ANA profile tests in children with autoimmune disorders; a retrospective study. Clin Rheumatol 2000;19:200–203. [DOI] [PubMed] [Google Scholar]
  • 23. Hauksdottιr GS, Jonsson T, Mellbye OJ. Antinuclear antibody (ANA) profile testing of ANA positive patients. Clin Rheumatol 2001;20:157–158. [DOI] [PubMed] [Google Scholar]
  • 24. Keech CL, Mccluskey J, Gordon TP. Transfection and overexpression of the human 60 Kda Ro/SS‐A autoantigen in Hep‐2 cells. Clin Immunol Immunopathol 1994;73:146–151. [DOI] [PubMed] [Google Scholar]
  • 25. Pourmand N, Blomberg S, Rönblomm L, Karlsson‐Parra A, Petterson I, Wahren‐ Herlenius M. Ro 52 kD autoantibodies are detected in a subset of ANA negative sera. Scand J Rheumatol 2000;29:116–123. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Laboratory Analysis are provided here courtesy of Wiley

RESOURCES