Accurate and prompt diagnosis of leptospiral infection in bovine is difficult due to limitation of current procedures. The present study describes the application of polymerase chain reaction (PCR) for detection of leptospiral deoxyribonucleic acid (DNA) in blood and urine samples in the spontaneous cases of bovine leptospirosis. In most of studies leptospires DNA detected by PCR in experimentally induced infection so in present study trying to evaluate this assay in naturally infected cattle and buffalo. A total of 120 blood and 304 urine samples were collected from cattle and buffaloes. These samples were subjected to PCR using two sets of primers i.e. G1/G2 and LipL32 and it revealed that 9/120 blood and 33/304 urine samples were found to be positive. The results suggested that PCR could be an excellent approach for diagnosis of Leptospirosis in field samples also. The specificity of primers was checked by using reference pathogenic serovar of Leptospira.

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