A zoonotic disease Leptospirosis is caused by pathogen of the genus Leptospira and it is an emerging global public health problem. In the present study a total of 266 cattle blood and sera samples were collected from the towns and villages of Nagpur, Wardha, Bhandara Gadchiroli and Durg districts during December 2017 to 2018 and tested for the presence of Leptospira. These samples were from randomly selected herds with history of repeated breeding, abortion, reproductive disorders, etc. also including some apparently healthy animals. The presence of leptospiral DNA in blood sample was assayed by PCR amplification of rrs (16S rRNA) gene. Antibodies against Leptospira serovars were tested using an enzyme-linked immunosorbent assay (ELISA) and microscopic agglutination test (MAT). Out of 266 blood samples, in 33 samples leptospires DNA was identified with the frequency of 12.40%. A total of 53 cattle sera were positive in commercial Leptospira Bovine Hardjo ELISA kit indicated 19.92% seroprevalence. In the MAT analysis 120 samples revealed presence of different serovars with the seropositivity 45.11%. The study supports the probable role of cattle in maintaining Leptospira Hardjo along with some other serovars and warrants an intensive control and surveillance programme for reducing leptospirosis in cattle.