Rapid diagnosis of leptospirosis is a major concern in many countries including Thailand. Unfortunately,the available conventional tests are time consuming, insufficiently sensitive and relatively tedious to perform. An effective and rapid detection assay is highly required. The present invention “LEP-Plus” is a sensitive, specific and rapid detection of acute infection by Leptospira spp. This lateral flow immunochromatographic assay employing patient’s serum or plasma to react with disease-specific lipopolysaccharide antigens immobilized on a strip of nitrocellulose is capable of detecting the elicited IgM antibody since early acute phase by 3 days of fever onset (1-5 days of illness). This allows early detection of leptospirosis and proper treatment given to prevent severe complications due to delayed diagnosis.