Leptospirosis is a disease which affects domestic and wild animals as well as humans. A highly conserved outer membrane lipoprotein LipL32 is considered to be an important antigen for detection of leptospirosis. In this study the recombinant protein LipL32 was extracted and characterized by immunochemical methods. The recombinant protein LipL32 was produced in Escherichia coli. We have optimized a LipL32 gene for expression in Rosetta (DE3) strain. 0.5% sodium lauryl sulfate was used to solubilize the protein and expressed into inclusion bodies. Immunochemical studies demonstrated a high reactivity of the recombinant LipL32 with antibodies elicited during infection of pathogenic Leptospira serovars. This study tested the diagnostic performance of the recombinant LipL32 in an in-house ELISA. The indirect ELISA was developed and its specificity was compared to that of a traditional microagglutination test (MAT).