In April 2014 foot-and-mouth disease virus (FMDV) affected water buffaloes (Bubalus bubalis) aged from 3-5 years in Qalyubia, Egypt. The aim of the present study was to diagnose FMDV molecularly and biochemically. Blood samples were collected from buffaloes suffering from characteristic clinical signs of FMDV infection as fever, profuse ruby threads salivation, ulcer on muzzle, vesicles on foot and lameness. Blood samples, tongue epithelium and vesicular fluid were evaluated by real time RT-qPCR for the diagnosis of FMDV using different probes and primers of universal (3D) gene and VP1 gene for serotypes A, Iran O, Asia and SAT2. The positive sample confirmed by one step reverse transcription polymerase chain reaction (RT-PCR). This resulted in the identification of a SAT2 serotype was the causative agent and the amplified RNA virus resulted in 716bp. Serum samples of positive PCR infected animals compared with apparently healthy control group was used to determine the concentration of aspartate amino transferase (AST), alanine amino transferase (ALT), alkaline phosphatase (ALP), albumin, total protein, calcium (Ca), iron (Fe) and inorganic phosphorus (Ph). A level of nitric oxide (NO) and malondialdehyde (MDA) were calorimetrically measured in serum as markers for oxidant status. There was a significant increase (P<0.05) in AST, ALT, ALP, Ph, NO and MDA and a significant decrease (P<0.05) in albumin, total protein, Ca and Fe in serum of clinically affected animals. It was concluded that FMDV serotype SAT2 circulate in Egypt and associated with biological alteration and disturbed oxidative status.