Numerous studies have been investigated to assess the relationship between the fertility related gene, CYP19, and reproduction in cow; however only scanty studies were performed on CYP19 gene in buffalo. The present study was designed to isolate a fragment of CYP19 gene containing exon 9 and to detect any polymorphism associated with infertility in Egyptian buffalo cows. CYP19 gene fragment was isolated by using PCR technique and detected by the direct sequencing and single strand conformation polymorphism (SSCP). A fragment (locus) of CYP19 gene with size of 446bp was successfully amplified. No SNP was detected in this locus as revealed by a monomorphic SSCP pattern observed in normal cyclic and anestrous animals. The results of SSCP were confirmed by nucleotide sequencing. Alignment of nucleotide sequences of this locus showed 100% identity with Indian water buffaloes (EF126034) and 99% identity with Bos Taurus (Z69249.1). The present study concluded that the sequence of CYP19 is highly conserved between cattle and buffalo.