Skip to content
Licensed Unlicensed Requires Authentication Published by De Gruyter September 21, 2011

Screening of antinuclear antibodies: comparison between enzyme immunoassay based on nuclear homogenates, purified or recombinant antigens and immunofluorescence assay

  • Sergio Bernardini , Maria Infantino , Lorenza Bellincampi , Marzia Nuccetelli , Antonella Afeltra , Roberta Iori , Antonino Biroccio , Andrea Urbani and Giorgio Federici

Abstract

Current clinical practice considers antinuclear antibody (ANA) testing as a screening test; this has a major impact on laboratory work with a growing volume of analyses that need to be performed rapidly, to maintain good specificity and sensitivity. Ongoing discussions have been raised in order to identify the best technology to use in ANA screening, taking into account both clinical and economical implications. The aim of our study was to compare three different enzyme immunoassays (EIA) with immunofluorescence (IF) assay in order to identify which test is better for use as a screening test. The study was performed on 473 sera and the three different EIA tests were based on nuclear homogenates from HeLa cells, purified antigens from HEp-2 cells and recombinant antigens, respectively. The concordance between EIA-ANA and IF-ANA techniques, determined by the κ statistic, was acceptable, but not complete, and discrepancies between both EIA-positive/IF-negative samples and IF-positive/EIA-negative were found. Both methods show interesting diagnostic abilities, however, the IF-ANA assay seems to be the first choice test in a well-standardized immunofluorescence laboratory with experienced microscopists, whereas the EIA test might be useful especially in large-scale ANA screening.

References

1 Kavanaugh A, Tomar R, Reveille R, Solomon DH, Hamburger HA. Guidelines for clinical use of the antinuclear antibody test for specific autoantibodies to nuclear antigens. Arch Pathol Lab Med 2000; 124:71–81.10.5858/2000-124-0071-GFCUOTSearch in Google Scholar

2 Gonzales C, Guevara P, Alarcon I, Hernando M, Navajo J, Gonzales-Buitrago JM. Antinuclear antibodies (ANA) screening by enzyme immunoassay with nuclear HEp-2 cell extract and recombinant antigens: analytical and clinical evaluation. Clin Biochem 2002; 35:463–9.10.1016/S0009-9120(02)00342-9Search in Google Scholar

3 Shmerling RH. Autoantibodies in systemic lupus erythematosus – there before you know it. N Engl J Med 2003; 349:1499–500.10.1056/NEJMp038142Search in Google Scholar PubMed

4 Arbuckle MR, McClain MT, Rubertone MV, Sconfield RH, Dennis GJ, James JA, et al. Development of autoantibodies before the clinical onset of systemic lupus erythematosus. N Engl J Med 2003; 349:1526–33.10.1056/NEJMoa021933Search in Google Scholar PubMed

5 Friou CJ. Clinical application of lupus serum nucleoprotein reaction using immunofluorescent antibody technique. J Clin Invest 1957; 36:890–7.Search in Google Scholar

6 Tan EM, Feltkamp TE, Smolen JS. Range of antinuclear antibodies in “healthy” individuals. Arthritis Rheum 1997; 40:1601–11.10.1002/art.1780400909Search in Google Scholar PubMed

7 Jaskowski TD, Schroder C, Martins TB, Mouristsen CL, Litwin CM, Hill HR. Screening for antinuclear antibodies by enzyme immunoassay. Am J Clin Pathol 1996; 106:468–73.10.1093/ajcp/105.4.468Search in Google Scholar PubMed

8 Homburger HA, Cahen YD, Griffiths J, Jacob GL. Detection of antinuclear antibodies: comparative evaluation of enzyme immunoassay and indirect immunofluorescence methods. Arch Pathol Lab Med 1998; 122:993–99.Search in Google Scholar

9 Emlen W, O’Neill L. Clinical significance of antinuclear antibodies: comparison of detection with immunofluorescence and enzyme-linked immunosorbent assays. Arthritis Rheum 1997; 40:1612–8.10.1002/art.1780400910Search in Google Scholar PubMed

10 Bayer PM, Bauerfeind S, Bienvenu J, Fabien N, Frei PC, Gilburd B, et al. Multicenter evaluation study on a new HEp2 ANA screening enzyme immune assay. J Autoimmun 1999; 13:89–93.10.1006/jaut.1999.0298Search in Google Scholar PubMed

11 Reisner BS, DiBlasi J, Goel N. Comparison of an enzyme immunoassay to an indirect fluorescent immunoassay for the detection of antinuclear antibodies. Am J Pathol 1999; 111:503–6.10.1093/ajcp/111.4.503Search in Google Scholar PubMed

12 Hayashi N, Kawamoto T, Mukai M, Morinobu A, Koshiba M, Kondo S, 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; 47:1649–59.10.1093/clinchem/47.9.1649Search in Google Scholar

13 Tan EM, Smolen JS, McDougal, Butcher BT, Conn D, Dawkins, et al. A critical evaluation of enzyme immunoassays for detection of antinuclear autoantibodies of denied specificities, I. Precision, sensitivity and specificity. Arthritis Rheum 1999; 42:455–64.10.1002/1529-0131(199904)42:3<455::AID-ANR10>3.0.CO;2-3Search in Google Scholar

14 Tan EM, Smolen JS, McDougal, Fritzler MJ, Gordon T, Hardin JA, et al. A critical evaluation of enzyme immunoassays for detection of antinuclear autoantibodies of denied specificities, II. Potential for quantification of antibody content. J Rheumatol 2002; 29:68–74.Search in Google Scholar

15 Koch GG, Landis JR, Freeman DH Jr, Lehnen RC. A general methodology for the analysis of experiments with repeated measurements of categorical data. Biometrics 1977; 33:133–58.10.2307/2529309Search in Google Scholar

16 Nossent H, Rekvig OP. Antinuclear antibody screening in this new millennium: farewell to the microscope? Scand J Rheumatol 2001; 30:123–6;127–8.10.1080/030097401300162860Search in Google Scholar

17 Feltkamp TEW. Antinuclear antibody determination in a routine laboratory. Ann Rheum Dis 1996; 55:723–7.10.1136/ard.55.10.723Search in Google Scholar

18 Genth E. Diagnostic evaluation of systemic autoimmune diseases. In: Peter HH, Pichler WJ, editors. Clinical immunology. Munich: Urban & Schwarzenberg, 1996: 119–33.Search in Google Scholar

19 Olaussen E, Rekvig OP. Screening tests for antinuclear antibodies (ANA): selective use of central nuclear antigens as a rationale basis for screening by ELISA. J Autoimmun 1999; 13:95–102.10.1006/jaut.1999.0295Search in Google Scholar

20 Miles J, Charles P, Riches P. A review of methods available for the identification of both organ-specific and non organ-specific autoantibodies. Ann Clin Biochem 1998; 35:19–47.10.1177/000456329803500104Search in Google Scholar

Received: 2004-7-9
Accepted: 2004-7-12
Published Online: 2011-9-21
Published in Print: 2004-10-1

© Walter de Gruyter

Downloaded on 26.5.2024 from https://www.degruyter.com/document/doi/10.1515/CCLM.2004.235/html
Scroll to top button