Article Text
Abstract
Introduction Visual evaluation of indirect immunofluorescence (IIF) on human epithelial-2 cells is the routine method for screening for antinuclear antibodies (ANA) in connective tissue diseases. Since visual IIF is time-consuming and subjective, automated IIF processors have been developed to offer standardised, valid and cost-efficient IIF assays.
Objective The aim of this study was to determine the diagnostic reliability of 2 widely used IIF processors (Aklides, Medipan GmbH and Helios, Aesku Diagnostics) under real-life laboratory working conditions.
Methods ANA were determined in samples from patients with suspected autoimmune rheumatic disease (n=1008) using both automated IIF processors and compared with the results obtained by visual interpretation. The performance of IIF processors to discriminate positive from negative samples, pattern recognition and end point titre prediction were evaluated.
Results The IIF processors showed moderate agreement with visual interpretation in discriminating positive from negative ANA samples (κ values: Aklides 0.494; Helios 0.415). The sensitivity/specificity was 89%/59% for Aklides and 87%/54% for Helios. However, both processors correctly identified 99% of definitely positive samples (titre ≥1:320). Aklides correctly identified 43% of fluorescence patterns and its light intensity values showed good correlation (Spearman's ρ=0.680) with visually obtained titres.
Conclusions Automated IIF determination under real-life laboratory working conditions remains a challenge. Owing to their high sensitivity at clinically relevant ANA titres, automated IIF processors can already support but not totally replace visual IIF.
- Autoimmunity
- Autoimmune Diseases
- Autoantibodies
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Footnotes
EF and G-RB contributed equally.
Contributors CDL carried out immunoassays, performed statistical analysis, contributed with patients data and drafted the manuscript. KE conceived of the study, carried out immunoassays, participated in its coordination and helped to draft the manuscript. EF conceived of the study, participated in its coordination, contributed with patients data and helped to draft the manuscript. G-RB conceived of the study, participated in its coordination, contributed with patients data and helped to draft the manuscript.
Funding BMBF (German ministry of education and science); ArthroMark 01EG1401A.
Competing interests None declared.
Patient consent Obtained.
Ethics approval All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and the 1964 Helsinki declaration and its later amendments or comparable ethical standards. The study was approved by the local ethics committee (EA/193/10).
Provenance and peer review Not commissioned; externally peer reviewed.
Data sharing statement No additional data are available.