Autoantibody detection using indirect immunofluorescence on HEp-2 cells

Ann N Y Acad Sci. 2009 Sep:1173:166-73. doi: 10.1111/j.1749-6632.2009.04735.x.


The detection of autoantibodies is an important element in the diagnosis and monitoring of disease progression in patients with autoimmune diseases. In laboratory diagnostic tests for connective tissue and autoimmune liver diseases, indirect immunofluorescence on HEp-2 cells plays a central role in a multistage diagnostic process. Despite the high quality of diagnostics, findings at different laboratories can differ considerably due to a lack of standardization, as well as subjective factors. The present paper formulates recommendations for the standardized processing and interpretation of the HEp-2 cell test for the detection of non-organ-specific (especially antinuclear) antibodies. It provides requirements regarding the diagnostic tests used, instructions for laboratory procedure and evaluation, and recommendations for interpretation. For an optimal laboratory diagnostic process, it is useful to have an informative, tentative clinical diagnosis and an experienced laboratory diagnostician. In addition, the following key elements are recommended: initial screening using indirect immunofluorescence on carefully chosen HEp-2 cells beginning with a serum dilution of 1:80 and evaluation under a microscope with powerful illumination; results from a titer of 1:160 upwards being considered positive; internal laboratory quality control; and standardized interpretation. The aim is to improve diagnostic tests and care of patients with autoimmune diseases as a central concern of the European Autoimmunity Standardization Initiative (EASI).

MeSH terms

  • Antibodies, Antinuclear / blood
  • Autoantibodies / blood*
  • Cell Line, Tumor
  • Fluorescent Antibody Technique, Indirect / methods*
  • Fluorescent Antibody Technique, Indirect / standards
  • Humans
  • Reference Standards
  • Reproducibility of Results


  • Antibodies, Antinuclear
  • Autoantibodies