The phylogenies based on nucleotide sequences using distance and neighbor-joining methods were congruent. The ITS-2 DNA region was amplified from Boophilus annulatus (Say, 1821) and Rhipicephalus sanguineus (Latreille, 1806) and was sequenced and analyzed to clarify the phylogenetic relationship of these Boophilus and Rhipicephalus species. The data suggested that the gene shared a similarity in sequence compared to ticks ITS-2 found in Gen Bank. The results indicated that the ITS-2 nucleotide sequence of B. annulatus strain Egypt and R. sanguineus strain Egypt reflected the phylogenetic relationship between the two species.