title: PCR-based molecular discrimination between Maritrema eroliae and Probolocoryphe uca (Digenea: Microphallidae) in Kuwait Bay.
abstract: Microphallid trematodes are common parasites in marine snails and crustacean hosts at Kuwait Bay. The larval stages of two microphallids, Maritrema eroliae and Probolocoryphe uca, are difficult to differentiate morphologically. In this study, two PCR-based techniques were established for quick and accurate discrimination between the larval stages of the two microphallid species, employing restriction fragment length polymorphism (PCR-RFLP) and species-specific primers. Both techniques utilized nucleotide differences in the second internal transcribed region (ITS2) of the ribosomal DNA (rDNA) in the two species. For the PCR-RFLP technique, restriction enzyme AvaII was selected and it generated different restriction profiles among the two microphallids. In addition, species-specific primers were prepared for each microphallid species that amplified distinctive fragments. Both techniques showed that the larval stages of the two microphallid species can be identified accurately. However, direct PCR amplification using species-specific primers was more advantageous than the PCR-RFLP technique since it allowed rapid and specific discrimination between the two species. This technique provides a useful tool that can be used in future studies for the study of the distribution of microphallid species and their definitive hosts at different localities of Kuwait Bay.
authors.affiliation: Department of Biological Sciences, Faculty of Science, Kuwait University.
authors.forename: W Y
authors.initials: WY
authors.lastname: Al-Kandari
authors.affiliation: Department of Biological Sciences, Faculty of Science, Kuwait University.
authors.forename: S A
authors.initials: SA
authors.lastname: Al-Bustan
authors.affiliation: Department of Biological Sciences, Faculty of Science, Kuwait University.
authors.forename: M
authors.initials: M
authors.lastname: Alnaqeeb
authors.affiliation: Department of Biological Sciences, Faculty of Science, Kuwait University.
authors.forename: A M
authors.initials: AM
authors.lastname: Isaac
mesh_terms: D000818:Animals; D017931:DNA Primers / Q000235:genetics; D018509:DNA, Helminth / Q000235:genetics*; D021903:DNA, Ribosomal Spacer / Q000235:genetics; D007730:Kuwait; D007814:Larva; D010274:Parasitology / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D014200:Trematoda / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification
chemical_list: D017931:DNA Primers; D018509:DNA, Helminth; D021903:DNA, Ribosomal Spacer