Fasciolosis in Dakhla Oasis, Elwadi El-Gadid, Egypt was investigated in its final host, cattle, and the intermediate host, Lymnaea trancatula snail. The study was conducted in 7 localities of Dakhla Oasis, Egypt. Fecal samples from 300 cattle were collected and examined, liver of slaughtered animals were inspected for flukes in abattoir. Moreover, Lymnaeidae snails were harvested from water streams and examined also. Flukes mitochondrial DNA-targeting PCR assay was carried out to amplify the extracted genomic DNA from both of infected snails and adult worms. F. hepatica eggs were found in 31.33% of examined cattle. Moreover, abattoir liver inspection revealed that 11.56% harbored adult flukes. The collected snails were identified as Lymnaea truncatula and Lymnaea cailliaudi. Furthermore, larval stages of F. hepatica (sporocyts, rediae and cercariae) were reported in 41/731 (5.61%) examined snails. However, L. cailliaudi snails were uninfetced. Using the conventional PCR, F. hepatica specific amplicon size of 1030 bp was detected in isolates obtained from different localities. Detection of F. hepatica in the infected snails with mitochondrial DNA-targeting PCR was validated with the specific amplicon size. Based on the presence of snail intermediate host and the conventional PCR findings, Fasciola species in Dakhla Oasis were identified as F. hepatica. Accordingly, zoonotic importance and hygienic measures must be taken in consideration and highlighted by veterinarians and authorized agencies in both urban and rural areas.