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The first recognizes only native dsDNA; the second recognizes both dsDNA and single-stranded DNA (ssDNA); the third reacts predominantly with denatured or ssDNA; and the fourth comprises anti-nucleosome autoantibodies, which recognize DNA-histone complexes and may yield results that differ from those obtained with assays specific for native dsDNA. Combining the Crithidia luciliae indirect immunofluorescence test with solid-phase immunoassays improves the diagnostic accuracy for anti-dsDNA antibody screening in patients with SLE. However, several limitations remain in the performance and interpretation of anti-dsDNA immassays, potentially leading to diagnostic errors. These pitfalls may arise mainly from analytical variability, assay design constraints, and a lack of standardized interpretation. Recent advances indicate a growing shift toward solid-phase multi-analyte arrays coupled with algorithm-based analysis, which may help reduce the seronegative gap in SLE while improving the harmonization of anti-dsDNA testing. 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