The present study was carried out for detection and identification of the isolated FMDV from infected tissue samples using serotyping antigen detection ELISA kit (IZSLER ELISA Kit).Thirty six lesion samples were collected from different localities (Menofia, Gharbia, Kalyoubia, Giza and Sharquia governorates) during the period from May 2014 to July 2015.The obtained results indicated that clinically, unvaccinated infected cattle in Gharbia governorate 2014 was positive to FMDV infection serotype O. Samples collected from infected, vaccinated cattle and buffaloes in Kalyoubia governorate during 2015 are positive to FMDV infection serotype O. Clinically infected cattle and buffaloes in Menofia governorate during 2015 seven samples were positive to FMDV infection, which serotyped as five samples were serotype O & two samples were SAT-2. Clinically infected cattle and buffaloes in Giza governorate during 2015 sixteen samples were positive to FMDV infection, which eight samples from them serotyped as SAT-2 & seven samples from them were serotype O & one of them was serotyped A. Clinically infected un vaccinated cattle in Sharquia governorate 2015 three samples were positive to FMDV infection serotype A. We concluded that large number of both vaccinated and un vaccinated animals in these governorates affected by FMDV infection by its different serotypes in the period of the study, and there are different factors affecting prevalence of FMDV as (age, state of vaccination, type of vaccination, sex, season and species).