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Updated: Apr 28, 2026
Anti-Nuclear Antibody Screening Using HEp-2 Cells
Published on: June 23, 2014
IV Brazilian guidelines for autoantibodies on HEp-2 cells
Paulo Luiz Carvalho Francescantonio1, Wilson de Melo Cruvinel1, Alessandra Dellavance2
1Pontifícia Universidade Católica de Goiás, Goiânia, GO, Brasil.
Insights
The Fourth Brazilian Consensus for Autoantibodies Screening in HEp-2 Cells (ANA) updated guidelines for indirect immunofluorescence assays. New patterns were added, emphasizing quality control and standardization for accurate autoantibody detection.
Area of Science:
- Immunology
- Cell Biology
- Medical Diagnostics
Background:
- Indirect immunofluorescence (IIF) on HEp-2 cells is a primary method for detecting antinuclear antibodies (ANA).
- Standardization and interpretation of ANA patterns are crucial for accurate autoimmune disease diagnosis.
- Previous consensus meetings have guided ANA testing in Brazil, but evolving techniques necessitate updates.
Purpose of the Study:
- To establish updated recommendations for ANA screening using HEp-2 cells in Brazil.
- To discuss strategies for technique, standardization, interpretation, and quality control of IIF assays.
- To address challenges in ANA pattern recognition and commercial kit variability.
Main Methods:
- Convened twenty-three experts in ANA testing from Brazilian academic and private laboratories.
- Facilitated discussions and consensus-building on recommendations for the fourth edition of the Brazilian ANA Consensus.
- Reviewed and agreed upon updates to the ANA testing algorithm and quality control measures.
Main Results:
- Incorporated three new patterns into the ANA algorithm: cytoplasmic Rods and Rings, nuclear Quasi-homogeneous, and CENP-F.
- Highlighted the complex mixed pattern associated with anti-DNA topoisomerase I (Scl-70) antibodies.
- Stressed the importance of continuous quality control, standardized screening dilutions, and conjugate titration.
Conclusions:
- The 4th ANA Consensus provides updated guidelines for ANA screening in Brazil.
- Emphasis on novel patterns, detailed description of complex patterns (e.g., Scl-70), and robust quality control is essential.
- Acknowledged commercial kit heterogeneity and recommended solid-phase methods for autoantibody confirmation.
Objective:
The Fourth Brazilian Consensus for Autoantibodies Screening in HEp-2 Cells (ANA) was held in Vitória, Espírito Santo, and aimed to discuss strategies and recommendations about the technique, standardization, interpretation and quality control of the indirect immunofluorescence reaction on HEp-2 cells.
Methods:
Twenty three ANA experts from university centers and private laboratories in different areas from Brazil discussed and agreed upon recommendations for the fourth edition of the Brazilian Consensus for Autoantibodies Screening in HEp-2 Cells.
Results And Conclusion:
The 4th ANA Consensus included three novel patterns into the existing algorithm (cytoplasmic Rods and Rings, nuclear Quasi-homogeneous, and CENP-F). Emphasis was given to the need of attention in describing the peculiar mixed pattern elicited by anti-DNA topoisomerase I (Scl-70) autoantibodies, comprising nuclear fine specked, nucleolar homogeneous pattern, NOR staining in metaphase plates, and cytoplasmic fine speckled patterns. The group also emphasized the need for continuous quality control in indirect immunofluorescence assays, the establishment of screening dilutions, as well as conjugate titration. An alert was made regarding the heterogeneity of commercial kits in defining patterns and the use of solid phase methodologies to determine the presence of autoantibodies.
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