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Updated: Jul 30, 2025

Anti-Nuclear Antibody Screening Using HEp-2 Cells
Published on: June 23, 2014
Laboratory and clinical practices in antinuclear antibody detection and related antigens: recommendations from a
Laura Martinez-Martinez1,2, Juan Irure-Ventura3,4, Aurora Jurado5
1Immunology Department, Hospital de La Santa Creu I Sant Pau, Universitat Autònoma de Barcelona, C/Sant Antoni M Claret 167, 08025, Barcelona, Spain. lmartinezma@santpau.cat.
Abstract:
Antinuclear antibodies (ANA) are the most widely used immunological test for the diagnosis of autoimmune diseases. Despite the recommendations of experts, there is some variability in performing and interpreting this test in routine practice. In this context, the Spanish Group on Autoimmune Diseases (GEAI) of the Spanish Society of Immunology (SEI) conducted a national survey of 50 autoimmunity laboratories. Here we report the survey results on ANA testing, detection of related antigens, and our recommendations. The survey showed that most of the participating laboratories use a similar approach for most key practices: 84% perform ANA by indirect immunofluorescence (IIF) on HEp-2 cells as the screening methodology while the other laboratories use IIF to confirm positive screens; 90% report ANA test results as either negative or positive with titer and pattern; 86% indicated that the ANA pattern conditioned follow-up testing for specific antigen-related antibodies; and 70% confirm positive anti-dsDNA. However, testing practices were highly heterogeneous for certain items, such as sera dilutions and the minimum time period for repeating ANA and related antigen determinations. Overall, this survey shows that most autoimmune laboratories in Spain use a similar approach but that further standardization of testing and reporting protocols is needed.
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