Introduction: The gold standard method to detect anti-nuclear antibodies is Indirect Immunofluorescence. Hep-2 cell lines coated slides are used to identify and detect the various patterns of fluorescence produced by anti-nuclear antibodies. Interpretation is based on the titre, pattern and intensity of fluorescence. Objectives: To find out whether any relationship exists between the intensity of fluorescence and the concentration of anti-nuclear antibodies. For this the samples which exhibited homogeneous pattern were further tested by quantitative Enzyme linked immunosorbent assay to find out the concentration of anti-dsDNA antibody. Materials and Methods: This prospective study was carried over a period of one year from January 2017 to December 2017 and the study group included 500 patients with lupus and 100 age and sex matched healthy controls. All the samples were processed for Indirect Immunofluorescence. The samples which were of homogeneous pattern were tested by Enzyme linked immunosorbent assay to quantitate the anti-dsDNA antibody concentration. Results: The anti-dsDNA positivity in homogeneous pattern was statistically significant (p Conclusion: As the intensity of fluorescence dose not reflect the concentration of antibody, further studies are needed to elucidate whether grading of intensity is mandatory in interpretation. Keywords: Anti-nuclear antibodies, Indirect Immunofluorescence, Intensity grading, Antibody concentration.