The study describes the first attempt to record leptospirosis in Erode by isolation and serological tests such as the microscopic agglutination test and IgM-based enzyme-linked immunosorbent assay. Twenty-nine clinically suspected cases showing fever, headache, body ache associated with jaundice, decreased urine output, and conjunctival suffusion were included. The age of the patients ranged between 10-71 years and most of them were agricultural workers. Paired sera were possible among 12 cases. All the patients had fever and headache and other more common symptoms were myalgia and icterus. Leptospiral culture was positive in 7 (24.1%) patients. Out of 29 patients, 26 (89.7%) were diagnosed as having current leptospiral infection based on serology and isolation. The leptospiral isolates KSR 1-6 were further characterized by using randomly amplified polymorphic DNA fingerprinting shown genetic similarities with Leptospira interrogans spp. This study shows the presence of leptospirosis among the hospital cases of Erode and that this disease is a potential health hazard of agricultural workers in Cauvery basin.