The evaluation of anti-dsDNA antibodies represents one of the essential diagnostic and prognostic marker features in patients affected with Systemic Lupus Erythematosus (SLE). In this study we have compared Immunoblotting (IB) with Crithidia luciliae Indirect Immunofluorescence test (CLIFT) and Chemiluminescent Immunoassay (CLIA) in 91 patients referred to our hospital for anti-dsDNA antibodies detection. The concordance and correlation measured by Cohen’s kappa and Spearman’s coefficient respectively was significant between CLIFT vs CLIA (0.70; 0,7404, p<0,0001) and among CLIA vs IB (0.79; 0,5377, p<0,0001) and lower between CLIFT vs IB (0.55; 0,4373, p>0,0001). Among the 46 IB positive samples, 14 were either positive for CLIA or CLIFT. It is noteworthy that 11 out of these 14 samples had the final diagnosis of SLE. Thirteen out of fourteen samples were also positive for anti-Nucleosome antibodies as measured concomitantly in Immunoblotting. While our observations are based on a limited number of samples and will have to be confirmed in a bigger cohort, they underline the contribution of immunoblotting as an additional assay in defining the anti-dsDNA antibody profile in association with other well established methods like CLIA and CLIFT.