The presence of ESBLs in many Gram negative strains are of serious concern, since these organisms are the most common cause of different human infections. ESBL positive phenotypically were tested for the presence of ESBL encoding genes using PCR with specific primers for the detection of CTX-M, TEM and SHV genes, then the amplicons were sequenced to characterized gene content. The presence of CTX-M, TEM and SHV genes was confirmed in 65 / 88 (73.9%) of the isolates. The ESBL genes were detected in 47 Escherichia coli, 14 Klebsiellapneumoniae, 2 Proteus mirabilis, 1 Serratiaodotifera and 1 Enterobactersakasaki. The nucleotide sequences were subjected to BLAST for sequences similarity and homology.