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AB1238 A comparison between automated and manual methods for detection antinuclear autoantibodies on human epithelial-2 cells
  1. A.A. Novikov1,
  2. E.N. Alexandrova1,
  3. D. Roggenbuck2,
  4. E.L. Nasonov3
  1. 1Laboratory of Immunology and Molecular Biology of Rheumatic Diseases, Fsbi “Institute of Rheumatology” Rams, Moscow, Russian Federation
  2. 2Medipan GmbH, Dahlewitz/Berlin, Germany
  3. 3Fsbi “Institute of Rheumatology” RAMS, Moscow, Russian Federation

Abstract

Background A variety of antinuclear antibodies (ANAs) are found in serum samples obtained from patients with rheumatic autoimmune diseases1. Identification of the corresponding antigens and measurement of the antibody titer are useful for diagnosis and to inform about the progress and prognosis of the disease2-4. The current gold standard method for ANA detection is manual indirect immunofluorescence (IIF) on human epithelial-2 (HEp-2) cells5.

Objectives Automation of ANAs IIF interpretation including pattern recognition can improve intra- and inter- laboratory assay reproducibility. Comparing automated and visual interpretation of ANAs the usefulness for laboratory diagnostics was investigated.

Methods Autoantibody detection by IIF on HEp-2 cells was performed in 234 serums of patients witch suspected and confirmed rheumatic disease. Automated evaluation (“Aklides platform”, Medipan GmbH and “ANA HEp-2 Plus” kit, Generic Assays GmbH) were compared with visual interpretations. The screening dilution was 1:160.

Results Automated method of recognition positive/negative results showed a moderate agreement with visual interpretation: kappa=0,5; case of mismatch is 30 (13%). The automated identification of the pattern (n=83, mixed patterns were excluded from this part of the study - this option is not available when using this automated platform) demonstrated a moderate agreement with visual assessment kappa=0,6; discrepant results: 22 (26,5%).

Conclusions Use of this automated platform for ANAs screening provides reliable identification of negative/positive samples and main nuclear patterns.

  1. Nakabayashi T, Kumagai T, PhD,Yamauchi K et al. Evaluation of the Automatic Fluorescent Image Analyzer, Image Titer, for Quantitative Analysis of Antinuclear Antibodies. Am J Clin Pathol 2001;115:424-429.

  2. Notman DD, Kurata N, Tan EM. Profiles of antinuclear antibodies in systemic rheumatic diseases. Ann Intern Med.1975; 83:464-469.

  3. Tan EM. Antinuclear antibodies in diagnosis and management. Hosp Pract (Off Ed). 1983;18:79-84.

  4. Tan EM. Antinuclear antibodies: diagnostic markers for autoimmune diseases and probes for cell biology. Adv Immunol. 1989;44:93-151.

  5. Kiviti S, Gilburd B, Agmon-Levin N et al. A novel automated indirect immunofluorescence autoantibody evaluation. Clin Rheumatol. 2011 Nov 5. [Epub ahead of print]

Disclosure of Interest None Declared

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