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The present study was included molecular study of three species that grouped into Hard ticks (Acari: Ixodidae) based on cytochrome oxidase I (COI) and 18S rRNA genes collected from three local areas' in An Najaf Province. Present results of study were showed that there were three abundant species of hard ticks identified as Boophilusannulatus, Rhipicephalussanguineus, and Hyalommadromedarii at prevalence ratio (48.1, 38.9 and 12.9) percentage respectively. Total number of hard ticks samples was (499) that infested on sheep and goat among them the females number were (228) more than males number were (155) whereas nymph number were lower (66). By using cytochrome oxidase I (COI) and 18S rRNA gene markers specific PCR assay present study was differentiated amongB. annulatus, R. sanguineus and H. dromedarii which abundant in An Najaf province. The results of gel electrophoresis based on specific PCR assay were detected that amplified segment size were 780 bp in18S rRNAgene and 700 bp in cytochrome oxidase I (COI) for species R. sanguineus and H.dromedarii respectively.