title: High-resolution melting analysis (HRM) for differentiation of four major Taeniidae species in dogs Taenia hydatigena, Taenia multiceps, Taenia ovis, and Echinococcus granulosus sensu stricto.
abstract: Tapeworms of the genus Taenia include several species of important parasites with considerable medical and veterinary significance. Accurate identification of these species in dogs is the prerequisite of any prevention and control program. Here, we have applied an efficient method for differentiating four major Taeniid species in dogs, i.e., Taenia hydatigena, T. multiceps, T. ovis, and Echinococcus granulosus sensu stricto. High-resolution melting (HRM) analysis is simpler, less expensive, and faster technique than conventional DNA-based assays and enables us to detect PCR amplicons in a closed system. Metacestode samples were collected from local abattoirs from sheep. All the isolates had already been identified by PCR-sequencing, and their sequence data were deposited in the GenBank. Real-time PCR coupled with HRM analysis targeting mitochondrial cox1 and ITS1 genes was used to differentiate taeniid species. Distinct melting curves were obtained from ITS1 region enabling accurate differentiation of three Taenia species and E. granulosus in dogs. The HRM curves of Taenia species and E .granulosus were clearly separated at Tm of 85 to 87 °C. In addition, double-pick melting curves were produced in mixed infections. Cox1 melting curves were not decisive enough to distinguish four taeniids. In this work, the efficiency of HRM analysis to differentiate four major taeniid species in dogs has been demonstrated using ITS1 gene.
authors.affiliation: Research Center for Hydatid Disease in Iran, Department of Parasitology, Kerman University of Medical Sciences, Kerman, 7616914115, Iran.
authors.forename: Mansoureh
authors.initials: M
authors.lastname: Dehghani
authors.affiliation: Research Center for Hydatid Disease in Iran, Department of Parasitology, Kerman University of Medical Sciences, Kerman, 7616914115, Iran.
authors.forename: Mohammad Ali
authors.initials: MA
authors.lastname: Mohammadi
authors.affiliation: Department of Research and Technology, Alborz University of Medical Sciences, Karaj, Iran.
authors.forename: Sima
authors.initials: S
authors.lastname: Rostami
authors.affiliation: Research Center for Hydatid Disease in Iran, Department of Parasitology, Kerman University of Medical Sciences, Kerman, 7616914115, Iran.
authors.forename: Saeedeh
authors.initials: S
authors.lastname: Shamsaddini
authors.affiliation: School of Medicine, Shahroud University of Medical Sciences, Shahroud, Iran.
authors.forename: Seyed Reza
authors.initials: SR
authors.lastname: Mirbadie
authors.affiliation: Research Center for Hydatid Disease in Iran, Department of Parasitology, Kerman University of Medical Sciences, Kerman, 7616914115, Iran. majid.fasihi@gmail.com.
authors.forename: Majid Fasihi
authors.initials: MF
authors.lastname: Harandi
mesh_terms: D000818:Animals; D051545:Cyclooxygenase 1 / Q000235:genetics; D018509:DNA, Helminth; D021901:DNA, Intergenic; D004283:Dog Diseases / Q000469:parasitology*; D004285:Dogs; D004443:Echinococcosis / Q000469:parasitology / Q000662:veterinary*; D048209:Echinococcus granulosus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D012756:Sheep / Q000469:parasitology; D013621:Taenia / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification
chemical_list: D018509:DNA, Helminth; D021901:DNA, Intergenic; D051545:Cyclooxygenase 1
keywords: Echinococcus granulosus; HRM analysis; ITS1; Melting curve; Taenia; Taeniidae