Abstract
Background: Automated systems for antinuclear antibody analysis are being introduced. The aim was to evaluate whether automated quantitative reading of fluorescence intensity is clinically relevant and allows for value-added reporting of test results.
Methods: Consecutive samples (n=260) were used to correlate fluorescence intensity with end-point titer. Moreover, 434 samples from controls (150 healthy blood donors, 150 chronic fatigue syndrome, and 134 diseased controls) and 252 samples (obtained at diagnosis) from patients with systemic rheumatic diseases were screened for antinuclear antibodies (1:80) on HEp-2 cells using NOVA View®, and likelihood ratios were calculated for fluorescence intensity result intervals.
Results: There was a significant correlation between end-point titer and fluorescence intensity. Likelihood ratios for a systemic rheumatic disease increased with increasing fluorescence intensity. The likelihood ratio for a systemic rheumatic disease was 0.06, 0.18, 0.51, 5.3, and 37.5 for a fluorescence intensity of ≤66, 67–150, 151–300, 301–1000, >1000, respectively. A range of 31%–37% of the patients with Sjögren’s syndrome, systemic sclerosis or systemic lupus erythematosus had fluorescence intensities >1000.
Conclusions: Estimation of fluorescence intensity by automated antinuclear antibody analysis offers clinically useful information. Likelihood ratios based on fluorescence intensity test result intervals aid with the interpretation of automated antinuclear antibody analysis and allow value-added reporting.
References
1. Solomon DH, Kavanaugh AJ, Schur PH; American College of Rheumatology Ad Hoc Committee on Immunologic Testing Guidelines. Evidence-based guidelines for the use of immunologic tests: antinuclear antibody testing. Arthritis Rheum 2002;47:434–44.10.1002/art.10561Search in Google Scholar PubMed
2. Op De Beeck K, Vermeersch P, Verschueren P, Westhovens R, Mariën G, Blockmans D, et al. Detection of antinuclear antibodies by indirect immunofluorescence and by solid phase assay. Autoimmun Rev 2011;10:801–8.10.1016/j.autrev.2011.06.005Search in Google Scholar PubMed
3. Op De Beéck K, Vermeersch P, Verschueren P, Westhovens R, Mariën G, Blockmans D, et al. Antinuclear antibody detection by automated multiplex immunoassay in untreated patients at the time of diagnosis. Autoimmun Rev 2012;12:134–43.Search in Google Scholar
4. Meroni PL, Schur PH. ANA screening: an old test with new recommendations. Ann Rheum Dis 2010;69: 1420–2.10.1136/ard.2009.127100Search in Google Scholar PubMed
5. Bizzarro N, Wiik A. Appropriateness in anti-nuclear antibody testing: from clinical request to strategic laboratory practice. Clin Exp Rheumatol 2004;22:349–55.Search in Google Scholar
6. Bossuyt X, Hendrickx A, Frans J. Antinuclear antibody titer and antibodies to extractable nuclear antigens. Arthritis Rheum 2005;53:987–8.10.1002/art.21602Search in Google Scholar PubMed
7. Tozzoli R, Bonaguri C, Melegari A, Antico A, Bassetti D, Bizzaro N. Current state of diagnostic technologies in the autoimmunology laboratory. Clin Chem Lab Med 2013;51: 129–38.10.1515/cclm-2012-0191Search in Google Scholar PubMed
8. Hiemann R, Büttner T, Krieger T, Roggenbuck D, Sack U, Conrad K. Challenges of automated screening and differentiation of non-organ specific autoantibodies on HEp-2 cells. Autoimmun Rev 2009;9:17–22.10.1016/j.autrev.2009.02.033Search in Google Scholar PubMed
9. Egerer K, Roggenbuck D, Hiemann R, Weyer MG, Büttner T, Radau B, et al. Automated evaluation of autoantibodies on human epithelial-2 cells as an approach to standardize cell-based immunofluorescence tests. Arthritis Res Ther 2010;12:R40.10.1186/ar2949Search in Google Scholar PubMed PubMed Central
10. Kivity S, Gilburd B, Agmon-Levin N, Carrasco MG, Tzafrir Y, Sofer Y, et al. A novel automated indirect immunofluorescence autoantibody evaluation. Clin Rheumatol 2012;31:503–9.10.1007/s10067-011-1884-1Search in Google Scholar PubMed
11. Voigt J, Krause C, Rohwäder E, Saschenbrecker S, Hahn M, Danckwardt M, et al. Automated indirect immunofluorescence evaluation of antinuclear antibodies on HEp-2 cells. Clin Dev Immunol 2012;2012:651058.10.1155/2012/651058Search in Google Scholar PubMed PubMed Central
12. Bossuyt X, Cooreman S, De Baere H, Verschueren P, Westhovens R, Blockmans D, et al. Detection of antinuclear antibodies by automated indirect immunofluorescence analysis. Clin Chim Acta 2012;415:101–6.10.1016/j.cca.2012.09.021Search in Google Scholar PubMed
13. Bonroy C, Verfaillie C, Smith V, Persijn L, De Witte E, De Keyser F, et al. Automated indirect immunofluorescence antinuclear antibody analysis is a standardized alternative for visual microscope interpretation. Clin Chem Lab Med 2013;51:1771–9.10.1515/cclm-2013-0016Search in Google Scholar PubMed
14. Tozzoli R, Antico A, Porcelli B, Bassetti D. Automation in indirect immunofluorescence testing: a new step in the evolution of the autoimmunology laboratory. Autoimmunity Highlights 2012;3:59–65.10.1007/s13317-012-0035-2Search in Google Scholar PubMed PubMed Central
15. Barak M, Rozenberg O, Grinberg M, Reginashvili D, Kishinewsky M, Henig C, et al. A novel cost effective algorithm for antinuclear antibody (ANA) testing in an outpatient setting. Clin Chem Lab Med 2013;51:e163–5.10.1515/cclm-2012-0535Search in Google Scholar PubMed
16. Fukuda K, Straus SE, Hickie I, Sharpe MC, Dobbins JG, Komaroff A. The chronic fatigue syndrome: a comprehensive approach to its definition and study. Ann Intern Med 1994;121:953–9.10.7326/0003-4819-121-12-199412150-00009Search in Google Scholar PubMed
17. Bossuyt X. Clinical performance characteristics of a laboratory test. A practical approach in the autoimmunity laboratory. Autoimmun Rev 2009;8:543–8.10.1016/j.autrev.2009.01.013Search in Google Scholar PubMed
18. American college of rheumatology ad hoc committee on immunologic testing guidelines. Guidelines for immunologic laboratory testing in the rheumatic diseases: an introduction. Arthritis Rheum 2002;27:429–33.10.1002/art.10381Search in Google Scholar PubMed
19. Vermeersch P, Bossuyt X. Comparative analysis of different approaches to report diagnostic accuracy. Arch Intern Med 2010;170:734–5.10.1001/archinternmed.2010.84Search in Google Scholar PubMed
©2014 by Walter de Gruyter Berlin/Boston
