Comparative Study of Human and Automated Screening for Antinuclear Antibodies by Immunofluorescence on HEp-2 Cells
DOI:
https://doi.org/10.6000/1929-6029.2015.04.03.4Keywords:
Autoantibodies, HEp-2, Immunofluorescence, autoimmune diseases, automated screening, standardizationAbstract
Background: Several automated systems had been developed in order to reduce inter-observer variability in indirect immunofluorescence (IIF) interpretation. We aimed to evaluate the performance of a processing system in antinuclear antibodies (ANA) screening on HEp-2 cells.
Patients and Methods: This study included 64 ANA-positive sera and 107 ANA-negative sera that underwent IIF on two commercial kits of HEp-2 cells (BioSystems® and Euroimmun®). IIF results were compared with a novel automated interpretation system, the “CyclopusCADImmuno®” (CAD).
Results: All ANA-positive sera images were recognized as positive by CAD (sensitivity = 100%), while 17 (15.9%) of the ANA-negative sera images were interpreted as positive (specificity = 84.1%), κ=0.799 (SD=0.045). Comparison of IIF pattern determination between human and CAD system revealed on HEp-2 (BioSystems®), a complete concordance in 6 (9.37%) sera, a partial concordance (sharing of at least 1 pattern) in 42 (65.6%) cases and in 16 (25%) sera the pattern interpretation was discordant. Similarly, on HEp-2 (Euroimmun®) the concordance in pattern interpretation was total in 5 (7.8%) sera, partial in 39 (60.9%) and absent in 20 (31.25%). For both tested HEp-2 cells kits agreement was enhanced for the most common patterns, homogenous, fine speckled and coarse speckled. While there was an issue in identification of nucleolar, dots and nuclear membranous patterns by CAD.
Conclusion: Assessment of ANA by IIF on HEp-2 cells using the automated interpretation system, the “CyclopusCADImmuno®” is a reliable method for positive/negative differentiation. Continuous integration of IIF images would improve the pattern identification by the CAD.
References
Wiik AS, Høier-Madsen M, Forslid J, Charles P, Meyrowitsch J. Antinuclear antibodies: A contemporary nomenclature using HEp-2 cells. J Autoimmun 2010; 35: 276-90. http://dx.doi.org/10.1016/j.jaut.2010.06.019 DOI: https://doi.org/10.1016/j.jaut.2010.06.019
Meroni PL, Schur PH. ANA screening: an old test with new recommendations. Ann Rheum Dis 2010; 69: 1420-2. http://dx.doi.org/10.1136/ard.2009.127100 DOI: https://doi.org/10.1136/ard.2009.127100
Bizzaro N, Antico A, Platzgummer S, Tonutti E, Bassetti D, Presente F, et al. Automated antinuclear immunofluore-scence antibody screening: A comparative study of six computer-aided diagnostic systems. Autoimmun Rev 2014; 13: 292-8. http://dx.doi.org/10.1016/j.autrev.2013.10.015 DOI: https://doi.org/10.1016/j.autrev.2013.10.015
Copple SS, Giles SR, Jaskowski TD, Gardiner AE, Wilson AM, Hill HR. Screening for IgG antinuclear antibodies by Hep-2 indirect immunofluorescence antibody assays and the need for standardization. Am J Clin Pathol 2012; 137: 825-30. http://dx.doi.org/10.1309/AJCPICNFG7UCES1S DOI: https://doi.org/10.1309/AJCPICNFG7UCES1S
Hahm D, Anderer U. Establishment of Hep-2 cell preparation for automated analysis of ANA fluorescence pattern. Cytometry A 2006; 69: 178-81. http://dx.doi.org/10.1002/cyto.a.20223 DOI: https://doi.org/10.1002/cyto.a.20223
Tonti S, Di Cataldo S, Bottino A, Ficarra E. An automated approach to the segmentation of Hep-2 cells for the indirect immunofluorescence ANA test. Comput Med Imaging Graph 2014. http://dx.doi.org/10.1016/j.compmedimag.2014.12.005b DOI: https://doi.org/10.1016/j.compmedimag.2014.12.005
Voigt J, Krause C, Rohwäder E, Saschenbrecker S, Hahn M, Danckwardt M, et al. Automated indirect immunofluore-scence evaluation of antinuclear autoantibodies on Hep-2 cells. Clin Dev Immunol 2012; 2012: 651058. http://dx.doi.org/10.1155/2012/651058 DOI: https://doi.org/10.1155/2012/651058
Egerer K, Roggenbuck D, Hienmann 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. http://dx.doi.org/10.1186/ar2949 DOI: https://doi.org/10.1186/ar2949
Hienmann 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. http://dx.doi.org/10.1016/j.autrev.2009.02.033 DOI: https://doi.org/10.1016/j.autrev.2009.02.033
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2015 Yousr Gorgi, Tarak Dhaouadi, Imen Sfar, Youssra Haouami, Taieb Ben Abdallah, Giuseppe Raso, Donato Cascio, Marco Cipolla, Vincenzo Taormina, Alessandro Fauci, Ignazio Brusca, Giuseppe Friscia, Amel Benammar Elgaaïed, Raja Marrakchi Triki, Asma Gati, Melika Ben Ahmed, Hechmi Louzir
This work is licensed under a Creative Commons Attribution 4.0 International License.
Policy for Journals/Articles with Open Access
Authors who publish with this journal agree to the following terms:
- Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
- Authors are permitted and encouraged to post links to their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work
Policy for Journals / Manuscript with Paid Access
Authors who publish with this journal agree to the following terms:
- Publisher retain copyright .
- Authors are permitted and encouraged to post links to their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work .
