Leptospirosis is emerging zoonotic infection in India with highest incidence in Gujarat, Maharashtra, Kerala & Karnataka. In human due to its varied clinical manifestation, diagnosis is difcult & relied mainly on serological test as cultural isolation & molecular diagnosis is tedious & costly. MAT is standard serological test for diagnosis. Present study was conducted on 238 blood samples of Leptospirosis suspected patients in South Gujarat region between years December 2013 to December 2015 in Department of Microbiology at New Civil Hospital, Surat after ethical approval. 16 positive isolates serum samples were subjected to the microscopic agglutination test (MAT). 07 (44%) patients were MAT positive out of 16 patients. Of the 07 MAT positive cases most pre-dominant serovars were australis and autumnalis with highest titre 1:200 and 1:400 respectively observed in 6 (38%) samples followed by icterohaemorrhagiae (highest titre 1:200 in 04 (25%) of the sample). For pomona and hebdomadis the highest titre was 1:100 and 1:50 respectively and was observed in 03 (19%) samples & grippotyphosa and pyrogens showed highest titre of 1:100 and 1:200 respectively and were observed in 02 (12%) samples. Patoc was the most predominant serogroup among non- pathogenic serovar giving the highest titre of 1:400 and was observed in 07 (44%) samples.