Abstract
IntroductionAnalysis of autoantibodies (AAB) by indirect immunofluorescence (IIF) is a basic tool for the serological diagnosis of systemic rheumatic disorders. Automation of autoantibody IIF reading including pattern recognition may improve intra- and inter-laboratory variability and meet the demand for cost-effective assessment of large numbers of samples. Comparing automated and visual interpretation, the usefulness for routine laboratory diagnostics was investigated.MethodsAutoantibody detection by IIF on human epithelial-2 (HEp-2) cells was conducted 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 results from routine diagnostics were compared with a novel automated interpretation system.ResultsBoth diagnostic procedures showed a very good agreement in detecting AAB (kappa = 0.828) and differentiating respective immunofluorescence patterns. Only 98 (8.0%) of 1222 sera demonstrated discrepant results in the differentiation of positive from negative samples. The contingency coefficients of chi-square statistics were 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. Comparing immunofluorescence patterns, 111 (15.3%) sera yielded differing results.ConclusionsAutomated assessment of AAB by IIF on HEp-2 cells using an automated interpretation system is a reliable and robust method for positive/negative differentiation. Employing novel mathematical algorithms, automated interpretation provides reproducible detection of specific immunofluorescence patterns on HEp-2 cells. Automated interpretation can reduce drawbacks of IIF for AAB detection in routine diagnostics providing more reliable data for clinicians.
Highlights
Analysis of autoantibodies (AAB) by indirect immunofluorescence (IIF) is a basic tool for the serological diagnosis of systemic rheumatic disorders
Due to the key position of anti-nuclear antibody (ANA) screening in the serological diagnosis of systemic rheumatic diseases, consistent reproducibility and high quality of human epithelial-2 (HEp-2) cell-based IIF assays are required [8,15,16]
ANAs were detected by HEp-2 cell assay and interpreted either visually by an experienced investigator or by automated reading and pattern recognition with the system
Summary
Analysis of autoantibodies (AAB) by indirect immunofluorescence (IIF) is a basic tool for the serological diagnosis of systemic rheumatic disorders. Automation of autoantibody IIF reading including pattern recognition may improve intra- and inter-laboratory variability and meet the demand for cost-effective assessment of large numbers of samples. Comparing automated and visual interpretation, the usefulness for routine laboratory diagnostics was investigated. Disease-specific autoantibodies (ABBs) are a serological phenomenon of systemic rheumatic conditions and autoimmune liver disorders. Due to the key position of ANA screening in the serological diagnosis of systemic rheumatic diseases, consistent reproducibility and high quality of HEp-2 cell-based IIF assays are required [8,15,16]. Automation of IIF pattern reading can provide a reliable basis for cost-effective serological diagnostics for laboratories with large sample numbers. The opportunity of modern electronic data management alleviates the heavy workload in such laboratories
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