Abstract
Indirect immunofluorescence (IIF) is the main technique for the detection of antinuclear antibodies (ANA) and antineutrophil cytoplasmic antibodies (ANCA). The fully automated IIF processor HELIOS® is the first IIF processor that is able to automatically prepare slides and perform automatic reading. The objective of the present study was to determine the diagnostic performance of this system for ANA and ANCA IIF interpretation, in comparison with visual IIF. ANA detection by visual IIF or HELIOS® was performed on 425 sera samples including: 218 consecutive samples submitted to a reference laboratory for routine ANA testing, 137 samples from healthy subjects and 70 ANA/ENA positive samples. For ANCA determination, 170 sera samples were collected: 40 samples for routine testing, 90 samples from healthy blood donors and 40 anti-PR3/anti-MPO positive subjects. Good correlation was found for the visual and automated ANA IIF approach regarding positive/negative discrimination of these samples (kappa = 0.633 for ANA positive samples and kappa = 0.657 for ANA negative samples, respectively). Positive/negative IIF ANCA discrimination by HELIOS® and visual IIF revealed a complete agreement of 100 % in sera from healthy patients and PR3/MPO positive samples (kappa = 1.00). There was 95 % agreement between the ANCA IIF performed by automated and visual IIF on the investigation of routine samples. Based on these results, HELIOS® demonstrated a high diagnostic performance for the automated ANA and ANCA IIF interpretation that was similar to a visual reading in all groups of samples.
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Abbreviations
- ANA:
-
Antinuclear antibodies
- ENA:
-
Extractable nuclear antigens
- ANCA:
-
Antineutrophil cytoplasmic antibodies
- cANCA:
-
Cytoplasmic ANCA
- pANCA:
-
Perinuclear ANCA
- MPO:
-
Myeloperoxidase
- PR3:
-
Proteinase 3
- ELISA:
-
Enzyme-linked immunosorbent assays
- IIF:
-
Indirect immunofluorescence
- SLE:
-
Systemic lupus erythematosus
- RA:
-
Rheumatoid arthritis
- SSc:
-
Systemic sclerosis
- IIM:
-
Idiopathic inflammatory myopathies
- SjS:
-
Sjögren’s syndrome
- HEp-2:
-
Human epidermoid laryngeal carcinoma
- ACR:
-
American College of Rheumatology
- AAV:
-
ANCA-associated vasculitis
References
Agmon-Levin N, Damoiseaux J, Shoenfeld Y. Response to: ‘detection of antinuclear antibodies—added-value of solid phase assay?’ by Bossuyt and Fieuws. Ann Rheum Dis. 2014;73:e11.
Fritzler MJ. Challenges to the use of autoantibodies as predictors of disease onset, diagnosis and outcomes. Autoimmun Rev. 2008;7:616–20.
Tozzoli R, Bizzaro N, Tonutti E, et al. Guidelines for the laboratory use of autoantibody tests in the diagnosis and monitoring of autoimmune rheumatic diseases. Am J Clin Pathol. 2002;117:316–24.
Meroni PL, Schur PH. ANA screening: an old test with new recommendations”. Ann Rheum Dis. 2010;69:1420–2.
Nifli AP, Notas G, Mamoulaki M, et al. Comparison of a multiplex, bead-based fluorescent assay and immunofluorescence methods for the detection of ANA and ANCA autoantibodies in human serum. J Immunol Methods. 2006;311:189–97.
Smith J, Onley D, Garey C, et al. Determination of ANA specificity using the UltraPlex platform. Ann NY Acad Sci. 2005;1050:286–94.
Invernizzi P. PBC screen: an IgG/IgA dual isotype ELISA detecting multiple mitochondrial and nuclear autoantibodies specific for primary biliary cirrhosis. J Autoimmun. 2010;35:436–42.
American College of Rheumatology, “American College of Rheumatology position statement: methodology of testing for antinuclear antibodies.” 2009. pp. 1–3.
Bosch X, Font J, Mirapeix E, et al. Anti-neutrophil cytoplasmic autoantibodies (ANCA): antigenic specificities and clinical associations. Adv Exp Med Biol. 1993;14:281–6.
Savige JF, Gillis DF, Benson E, et al. International consensus statement on testing and reporting of antineutrophil cytoplasmic antibodies (ANCA). Am J Clin Pathol. 1999;14:507–13.
Craig WY, Ledue TB, Collins MF, et al. Serologic associations of anti-cytoplasmic antibodies identified during anti-nuclear antibody testing. Clin Chem Lab Med. 2006;14:1283–6.
Csernok E, Ahlquist D, Ullrich S, et al. A critical evaluation of commercial immunoassays for antineutrophil cytoplasmic antibodies directed against proteinase 3 and myeloperoxidase in Wegener’s granulomatosis and microscopic polyangiitis. Rheumatology (Oxford). 2002;14:1313–7.
Egerer K, Roggenbuck D, Hiemann R 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.
Hiemann R, Hilger N, Michel J, et al. Automatic analysis of immunofluorescencepatterns of HEp-2 cells. Ann NY Acad Sci. 2007;1109:358–71.
Bonroy C, Verfaillie C, Smith V, et al. Automated indirect immunofluorescence antinuclear antibody analysis is a standardized alternative for visual microscope interpretation. Clin Chem Lab Med. 2013;51(9):1771–9.
Voigt J, Krause C, Rohwäder E, et al. Automated indirect immunofluorescence evaluation of antinuclear autoantibodies on HEp-2 cells. Clin Dev Immunol. 2012;2012:651058.
Knütter I, Hiemann R, Brumma T, et al. Automated interpretation of ANCA patterns: a new approach in the serology of ANCA-associated vasculitis. Arthritis Res Ther. 2012;14(14):R271.
Kivity S, Gilburd B, Agmon-Levin N, et al. A novel automated indirect immunofluorescence autoantibody evaluation. Clin Rheumatol. 2011;14:503–9.
Willitzki A, Hiemann R, Peters V, et al. New platform technology for comprehensive serological diagnostics of autoimmune diseases. Clin Dev Immunol. 2012;2012:284740.
Melegari A, Bonaguri C, Russo A, et al. A comparative study on the reliability of an automated system for the evaluation of cell-based indirect immunofluorescence. Autoimmun Rev. 2012;11:713–6.
Tozzoli R, Bonaguri C, Melegari A, et al. Current state of diagnostic technologies in the autoimmunology laboratory. Clin Chem Lab Med. 2013;51:129–38.
Tozzoli R, Antico A, Porcelli B, et al. Automation in indirect immunofluorescence testing: a new step in the evolution of the autoimmunology laboratory. Autoimmun Highlight. 2012;2012(3):59–65.
Bizzaro N, Antico A, Platzgummer S, et al. Automated antinuclear immunofluorescence antibody screening: a comparative study of six computer-aided diagnostic systems. Autoimmun Rev. 2014;13(3):292–8.
Damoiseaux J, Mallet K, Vaessen M, et al. Automatic reading of ANCA-slides: evaluation of the AKLIDES system. In: Conrad K, Chan EKL, Fritzler MJ, Humbel RL, Meroni PL, Shoenfeld Y, editors. From prediction to prevention of autoimmune disease. Lengerich, Germany: Pabst Science Publishers; 2011. p. 683–4.
http://www.aesku-systems.com/index.php/systems-ifa/helios-ifa-processor.
Altman D. Practical statistics for medical research. London, UK: Chapman & Hall; 1991.
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Shovman O and Agmon-Levin N have contributed equally to this work.
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Shovman, O., Agmon-Levin, N., Gilburd, B. et al. A fully automated IIF system for the detection of antinuclear antibodies and antineutrophil cytoplasmic antibodies. Immunol Res 61, 135–140 (2015). https://doi.org/10.1007/s12026-014-8588-y
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DOI: https://doi.org/10.1007/s12026-014-8588-y