Abstract
Indirect immunofluorescence antinuclear antibodies (IIF-ANA) are detected in approximately 90% of scleroderma patients, and the staining pattern correlates with scleroderma-specific antibody subsets. Solid-phase ANA assays that are dependent on multiplex bead technology (MULTIPLEX-ANA) are replacing immunofluorescence in many commercial labs; however, performance of these assays has not been compared to IIF-ANA in scleroderma. The purpose of this study was to evaluate whether a proportion of scleroderma patients have negative testing on MULTIPLEX-ANA assays and demonstrate whether negative MULTIPLEX-ANA is associated with particular scleroderma-specific autoantibodies. A retrospective chart review was completed on all 238 scleroderma patients evaluated in the Georgetown scleroderma clinic between June 1, 2008 and May 31, 2009. Autoantibody results, demographics, and scleroderma features were collected. Data were analyzed using unpaired t test and Mann–Whitney U test for continuous variables, and Fisher’s exact test for dichotomous variables. Simple kappa coefficient was used to measure the level of agreement between MULTIPLEX-ANA and IIF-ANA results. Two-tailed p values <0.05 were considered significant. MULTIPLEX-ANA testing was available in 57 patients and only 29 (51%) tested positive. In contrast, IIF-ANA was positive in 91% of these patients. Using simple kappa coefficient, there was a good agreement between the MULTIPLEX-ANA, and presence of Scl70, RNP, and centromere antibodies (0.76; 95% CI 0.59, 0.92), but there was no agreement between MULTIPLEX-ANA and presence of other IIF-ANA patterns including nucleolar ANA (−0.40; 95% CI −0.64, −0.16). Because RNA polymerase III and nucleolar antibodies are seen in 43% of the entire scleroderma population, we are concerned that these false-negative tests could result in delays in referral and diagnosis. Until the MULTIPLEX-ANA assays can be modified to include the antigens for RNA polymerase III and the nucleolar ANA subsets, IIF-ANA remains the recommended screening test for ANA in suspected scleroderma.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Lyons R, Narain S, Nichols C, Satoh M, Reeves WH (2005) Effective use of autoantibody tests in the diagnosis of systemic autoimmune disease. Ann N Y Acad Sci 1050(1):217–228. doi:10.1196/annals.1313.023
Steen VD (2005) Autoantibodies in systemic sclerosis. Semin Arthritis Rheum 35(1):35–42
Koenig M, Dieudé M, Senécal J-L (2008) Predictive value of antinuclear autoantibodies: the lessons of the systemic sclerosis autoantibodies. Autoimmun Rev 7(8):588–593. doi:10.1016/j.autrev.2008.06.010
Eissfeller P, Sticherling M, Scholz D, Hennig K, Lüttich T, Motz M, Kromminga A (2005) Comparison of different test systems for simultaneous autoantibody detection in connective tissue diseases. Ann N Y Acad Sci 1050(1):327–339. doi:10.1196/annals.1313.035
Shovman O, Gilburd B, Barzilai O, Shinar E, Larida B, Zandman-Goddard G, Binder SR, Shoenfeld Y (2005) Evaluation of the BioPlex™ 2200 ANA screen: analysis of 510 healthy subjects: incidence of natural/predictive autoantibodies. Ann N Y Acad Sci 1050(1):380–388. doi:10.1196/annals.1313.120
Shovman O, Gilburd B, Zandman-Goddard G, Yehiely A, Langevitz P, Shoenfeld Y (2005) Multiplexed AtheNA multi-lyte immunoassay for ANA screening in autoimmune diseases. Autoimmunity 38(1):105–109. doi:10.1080/08916930400022707
ACR (2009) Methodology of testing for antinuclear antibodies. http://www.rheumatology.org/practice/ana_position_stmt.pdf. Accessed March 2011.
Hanly JG, Su L, Farewell V, Fritzler MJ (2010) Comparison between multiplex assays for autoantibody detection in systemic lupus erythematosus. J Immunol Meth 358(1–2):75–80. doi:10.1016/j.jim.2010.04.005
Copple SS, Martins TB, Masterson C, Joly E, Hill HR (2007) Comparison of three multiplex immunoassays for detection of antibodies to extractable nuclear antibodies using clinically defined sera. Ann N Y Acad Sci 1109(1):464–472. doi:10.1196/annals.1398.052
Martins TB, Burlingame R, von Muhlen CA, Jaskowski TD, Litwin CM, Hill HR (2004) Evaluation of multiplexed fluorescent microsphere immunoassay for detection of autoantibodies to nuclear antigens. Clin Diagn Lab Immunol 11(6):1054–1059. doi:10.1128/cdli.11.6.1054-1059.2004
LeRoy EC, Medsger TA (2001) Criteria for the classification of early systemic sclerosis. J Rheumatol 28(7):1573–1576
Okano Y (1996) Antinuclear antibody in systemic sclerosis (scleroderma). Rheum Dis Clin North Am 22(4):709–735
Hamaguchi Y (2010) Autoantibody profiles in systemic sclerosis: predictive value for clinical evaluation and prognosis. J Dermatol 37(1):42–53. doi:10.1111/j.1346-8138.2009.00762.x
Nguyen B, Assassi S, Arnett FC, Mayes MD (2010) Association of RNA polymerase III antibodies with scleroderma renal crisis. J Rheumatol 37(5):1068. doi:10.3899/jrheum.091048
Parker J, Burlingame R, Webb T, Bunn C (2008) Anti-RNA polymerase III antibodies in patients with systemic sclerosis detected by indirect immunofluorescence and ELISA. Rheumatology 2008(47):976–979
Kuwana M, Okano Y, Pandey JP, Silver RM, Fertig N, Medsger TA (2005) Enzyme-linked immunosorbent assay for detection of anti-RNA polymerase III antibody: analytical accuracy and clinical associations in systemic sclerosis. Arthritis Rheum 52(8):2425–2432. doi:10.1002/art.21232
Acknowledgments
Dr. Shanmugam’s work is supported by award number KL2RR031974 and UL1RR031975 from the National Center for Research Resources.
Disclosures
None.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Shanmugam, V.K., Swistowski, D.R., Saddic, N. et al. Comparison of indirect immunofluorescence and multiplex antinuclear antibody screening in systemic sclerosis. Clin Rheumatol 30, 1363–1368 (2011). https://doi.org/10.1007/s10067-011-1766-6
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10067-011-1766-6