Aim: This study was undertaken to estimate the seroprevalence of leptospirosis among subjects with fever and to compare three diagnostic methods used in leptospirosis among Chennai population. Materials and methodology: Two hundred and sixty seven patients were included in the study. All the cases had history of fever for five or more days of the total study entrants, all were tested for Leptospira antibodies by MSAT, ELISA as well as MAT and the antigen detection was done using Dark Field Microscopy. The common clinical conditions with which the patients presented were 267 cases of fever, 89 cases of myalgia, headache in 63 cases, jaundice in 10 cases, conjunctival suffusion in 9 cases, diarrhea in 3 cases and nausea/vomiting in 35 cases. Results: Comparison of efficacy of MSAT and MAT was done. 92 cases were positive on Microscopic agglutination test (MAT) and 116 were positive on MSAT. All the MAT positive cases were positive on DFM. Though DFM alone is not a reliable test, it was compared with gold standard ELISA test and the findings shows that all DFM positive cases were also positive for MAT. However the number of positive cases detected using MSAT was higher than that of MAT. On comparing the results of MSAT and ELISA with MAT, there was 82% agreement between the tests.