Arthritis & Rheumatism, Volume 62,
November 2010 Abstract Supplement

Abstracts of the American College of
Rheumatology/Association of Rheumatology Health Professionals
Annual Scientific Meeting
Atlanta, Georgia November 6-11, 2010.

Standardization of Antinuclear Antibody Assessment on HEp-2 Cells by Automated Interpretation.

Egerer1,  Karl, Roggenbuck4,  Dirk, Hiemann6,  Rico, Weyer5,  Max-Georg, Lehmann3,  Barbara, Feist3,  Eugen, Burmester2,  Gerd R.

Charite-University Medicine, Berlin, Germany
Charite-University Medicine, Berlin, Germany
Charite-University Medicine
Medipan GmbH, Berlin Dahlewitz, Germany
Medizinisches Versorgungszentrum für Laboratoriumsmedizin, Mikrobiologie, Virologie und Infektionsepidemiologie, Hygiene und Umweltmedizin
University of Applied Science Lausitz


Analysis of antinuclear antibodies (ANAs) by indirect immunofluorescence (IIF) is a basic tool for the serological diagnosis of systemic rheumatic disorders. Automation of ANA IIF interpretation including pattern recognition can improve assay reproducibility and intra- and inter-laboratory variability. Automation of ANA reading meets the demand for cost-effective assessment of large numbers of samples in routine laboratories. Comparing automated and visual interpretation of ANA patterns, the usefulness for laboratory diagnostics was investigated.


Antinuclear antibody detection by IIF on human epithelial-2 (HEp-2) cells including pattern recognition was performed in a total of 1222 consecutive sera of patients with suspected systemic rheumatic diseases from a university routine laboratory (n=924) and a private referral laboratory (n=298). IIF reading results obtained in routine diagnostics were compared with findings by a novel automated interpretation system employing mathematical pattern recognition algorithms.


Visual and automated interpretation of ANA showed a very good agreement regarding positive / negative discrimination (kappa=0.828). Only 98 (8.0%) of 1222 sera demonstrated discrepant results in the differentiation of positive from negative samples. The contingency coefficient of Chi-square statistics was 0.646 for the university laboratory cohort with an agreement of 93.0% and 0.695 for the private laboratory cohort with an agreement of 90.6%, p<0.0001, respectively. According to the McNemar test, there was no significant difference in the university cohort (1.08%, p=0.25).

Comparing respective immunofluorescence patterns, 111 (15.3%) sera yielded differing results in both cohorts. Discrepant results were mainly obtained when the AKLIDES algorithms assessed cytoplasmic signals as nuclear staining due to superposition.


Automated interpretation of ANA by IIF on HEp-2 cells using an automated reading system is a reliable and robust method for positive/negative differentiation of IIF findings. Employing novel mathematical algorithms, automated interpretation provides reproducible detection of specific immunofluorescence patterns on HEp-2 cells. Automated interpretation can reduce drawbacks of ANA detection by IIF in routine diagnostics providing more reliable data for clinicians.

To cite this abstract, please use the following information:
Egerer, Karl, Roggenbuck, Dirk, Hiemann, Rico, Weyer, Max-Georg, Lehmann, Barbara, Feist, Eugen, et al; Standardization of Antinuclear Antibody Assessment on HEp-2 Cells by Automated Interpretation. [abstract]. Arthritis Rheum 2010;62 Suppl 10 :911
DOI: 10.1002/art.28679

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