Out of 100 milk samples collected from 45 buffaloes suffering from clinical mastitis and 55 milk samples collected from 19 apparent healthy buffaloes, showed that 7 samples were positive for Mycoplasma from mastitic cases meanwhile no Mycoplasma was obtained culturally from apparent healthy cases. PCR and indirect enzyme linked immunosorbent assay (ELISA) were used here in this study as a modern technique for diagnosis and results obtained showed that all isolates with the goover samples proved by conventional methods to be Mycoplasma bovis were also found to be positive by the two methods. Clinicopathological picture was described by haematological and biochemical examination of blood and serum ‘samples of apparent ne healthy andmastitic buffaloes.