Application of RAPD-PCR Technique in Molecular Genetic Approach for Detection of Genetic Variability

Understanding genetic structure and status of genetic variation of the Fasciola gigantica and F. hepatica isolates from different hosts, has important implications for epidemiology and effective control of fasciolosis. The aim of the present work was to study the molecular characterization of Fasciola gigantica and F. hepatica isolates collected from cows and sheep, using the random amplified polymorphic DNA fragments-polymerase chain reaction (RAPDsPCR) technique. Methods: F. gigantica and F. hepatica of bovine and ovine origin were collected from the biliary tracts and gall bladders of condemned bovine and ovine livers from Cairo Governmental slaughter house. By using (RAPDs-PCR) technique, optimal standardization of conditions of amplification and thermocyclation was made, using genetic markers. The methodology used compared the genetic pattern between the two species (inter-species) and inside each species (intra-species) between cow and sheep and the amplification fragments were between 135 and 741 base pairs of marker. Results: The results showed genetic variations (polymorphisms) of Fasciola gigantica and F. hepatica with amplification fragment based on a 500 – 400 base pair (bp). Inside each species, there were genetic variations in bovine and ovine strains and the amplification fragments were between 600 and 400 base pairs (bp). Conclusion: This assay is useful for both individual diagnosis and epidemiological surveys in endemic regions. [New York Science Journal 2010;3(11):52-60]. (ISSN: 1554-0200).