Escherichia coli is considered as one of the bacteria that causing diarrhea outbreaks all over the world, and it is responsible for diseases for human and animals as well A total number of 50 raw milk and 50 raw beef meat samples were collected from the local market in Fayoum Governorate. These samples were subjected for bacteriological, serological and molecular investigations. E. coli was isolated from raw milk and raw beef samples with an isolation rate of 58% and 14%, respectively. Serogrouping of the E. coli isolates from the raw milk samples revealed presence of O142, O55, O111, O27, and O26 with percentage of 20.69%, 20.69%, 17.24%, 17.24%, and 3.45%, respectively. However, the serogrouping of the E. coli isolates from raw beef meat samples revealed presence of O111, O27, O142, O55, and O127 with percentage of 28.56%, 14.28%, 14.28%, 14.28%, and 14.28%, respectively. Multiplex PCR was applied for the detection of virulence genes including shiga-toxin genes (stx1 and stx2), and the intimin gene (eae) which detected in E. coli. All the isolates were negative to both stx1 and stx2 genes. Meanwhile, the raw milk isolates of O142, O55, O111, and O27 were positive to eae gene. However, O26 isolate was negative to this gene. Also, the raw beef meat isolates of O142, O55, O111, and O27 were positive to eae gene. But, O127 isolate was negative to this gene.