One of the neglected food -born-diseases is fascioliasis ,it is serious parasitic disease infecting humans and animals worldwide and tops all the zoonotic helminthes .In Egypt , human and animal fasciolisis is an endemic clinical and epidemiological health problem .The causative agents, Fasciola hepatica and Fasciola gigantica, which their geographical distributions is overlapped. Moreover, in humans, liver flukes classification can not be achieved by clinical , pathological or immunological method. The differential diagnosis between Fasciola hepatica and Fasciola gigantica
infection is very important because of their different transmission and epidemiological characteristics . A simple and rapid PCR-restriction fragment length polymorphism (RFLP) assay, using the common restriction enzyme Avail is described to distinguish between both fascioloid species.
In our study , adults of F. hepatica and F. gigantica were isolated from liver of naturally infected sheep and cattle brought from various regions of Sohag Governorate. The 2 species were identified based on morphological criteria and all doubtful samples were discare and (QIA amp DNA Mini Kit) used, a new method to isolate DNA from the worm and Restriction fragment length polymorphism patterns of worms were obtained after
digestion of the PCR products with Avail restriction enzymes. The result of a regular PCR experiment for the amplification of the selected 28S rDNA fragment with the designed primer set yielded identical 648- bp-long PCR
products for F. hepatica and F. gigantica. Where the RFLP profile obtained From F. hepatica revealed three fragments of 628, 358 and 300 bp, and F. gigantica generated a clearly distinguishable four fragments corresponding to 628, 541 , 358 and 300 bp fragments. This results referred that two species present in our locality ,either F.gigantica and F. hepatica as there are different band patterns generated after digestion or presence of intermediate forms as there are three similar fragments.
This assay is a simple , rapid and reliable method for differentiating Fasciola spp,and can be used for diagnostic and epidemiological purposes.