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Differential diagnosis of Egyptian Fasciola species by deoxyribonucleic acid (DNA) sequences of nuclear ribosomal internal transcribed spacer 1 (ITS1) based on a PCR-RFLP method

Fascioliasis is one of the familiar zoonotic health problems of worldwide distribution including Asian, African countries and Egypt. Morphological differentiation between the liver fluke species Fasciola hepatica and Fasciola gigantica is difficult. Molecular characterization is important for discriminating Fasciola specimens having the deoxyribonucleic acid (DNA) sequences of Fasciola hepatica, Fasciola gigantica. We have developed a restriction fragment length polymorphism of amplified DNA (PCR-RFLP) of the nuclear ribosomal internal transcribed spacer 1 (ITS1) region in Fasciola species. The band patterns of the fragments digested with a restriction enzyme, Rsa I, were accurately distinguished between both forms of Fasciola. Amplicons with the sequences of F. hepatica and F. gigantica were divided into fragments of about 360, 100, and 60 bp, and 360, 170, and 60 bp, respectively, and amplicons with the sequences of both Fasciola species yielded fragments of 360, 170, 100, and 60 bp. The results of PCR-RFLP completely coincided with those of sequence analysis, and thus PCR-RFLP is a useful technique for determining the ITS1 type in Fasciola species. (Khaled Abd El-Aziz Mohammad, Alaa Abd El-Aziz Mohammad, Mohammad Fathallah Abu El-Nour and Anwar Husainy Abu-Hashim. Differential Diagnosis of Egyptian Fasciola species by deoxyribonucleic acid (DNA) sequences of nuclear ribosomal internal transcribed spacer 1 (ITS1) based on a PCR-RFLP method. N Y Sci J. 2012;5(5):46-49). (ISSN: 1554-0200). http://www.sciencepub.net/newyork . 9