In the present study, we report the isolation of BVDV field strains from apparently health buffaloes. One hundred and thirty-two samples including buffy coat and organs were collected from apparently health Screening buffaloes in Cairo, Mansoura and Suez governorates in Egypt during the year 1999. Screening of samples by GAPT using standard known anti-BVDV antisera revealed that three positive samples for the presence of BVDV antigen. In a trial for the isolation of the BVDV, 69 buffy coat samples were inoculated in MDBK cells, five selected samples that induced a clear cytopathic effect on inoculated cells were passaged for 7 times and subjected for further antigenic and molecular characterization using virus neutralization test and different formats Of PCR assays. The results revealed that the isolated BVDV field strains (3 from Cairo designated as Cairo/10/99: Cairo/12/99 and Cairo/13/99 and 2 from Suez designated as Suez/60/99 and Suez/68/99) were of BVDV type I.