This study aims to determine the SAT-2 serotype causing FMD outbreak in buffalo in Wassit and Dhi-Qar governorates, Iraq. This was done by clinical and serological methods. ELISA technology, which measures antibody levels in buffalo serum samples. 92 blood samples were collected from infected buffalo for antibody screening against FMDV serotype SAT-2 in infected animals which develop antibodies against the non-structural proteins (NSP) of the virus. Affected animals showed various clinical signs, such as fever, decreased appetite, mouth and foot vesicles, and lameness. According to the results of serological tests, the infection rate reached 85/92 positive samples (92.39%). The results showed, Age and sexspecific investigations of FMDV seropositivity did not reveal any statistically significant differences. The age groups (6-18 months and 18-36 months) had lowest infection rate (91.42%) and (92%) respectively, where as the age group 36 months, had highest rate of infection at (93.75%). Also, the results revealed, the seroprevalence rates infection for males were (90.6%), which had lowest than females (93.33%). This study, confirms a widespread outbreak of FMD caused by the SAT2 serotype in buffalo populations across the Wassit and Dhi-Qar governorates of Iraq. The study's findings may offer helpful information about how FMD protection measures are established and carried out in Iraq.