title: A dual PCR-based sequencing approach for the identification and discrimination of Echinococcus and Taenia taxa.
abstract: Reliable and rapid molecular tools for the genetic identification and differentiation of Echinococcus species and/or genotypes are crucial for studying spatial and temporal transmission dynamics. Here, we describe a novel dual PCR targeting regions in the small (rrnS) and large (rrnL) subunits of mitochondrial ribosomal RNA (rRNA) genes, which enables (i) the specific identification of species and genotypes of Echinococcus (rrnS + L-PCR) and/or (ii) the identification of a range of taeniid cestodes, including different species of Echinococcus, Taenia and some others (17 species of diphyllidean helminths). This dual PCR approach was highly sensitive, with an analytical detection limit of 1 pg for genomic DNA of Echinococcus. Using concatenated sequence data derived from the two gene markers (1225 bp), we identified five unique and geographically informative single nucleotide polymorphisms (SNPs) that allowed genotypes (G1 and G3) of Echinococcus granulosus sensu stricto to be distinguished, and 25 SNPs that allowed differentiation within Echinococcus canadensis (G6/7/8/10). In conclusion, we propose that this dual PCR-based sequencing approach can be used for molecular epidemiological studies of Echinococcus and other taeniid cestodes.
authors.affiliation: Institute of Parasitology, University of Bern, Bern, Switzerland; University of Monastir, Faculty of Pharmacy, Department of Clinical Biology B, Laboratory of Medical and Molecular Parasitology-Mycology (LR12ES08), Monastir, Tunisia.
authors.forename: Ghalia
authors.initials: G
authors.lastname: Boubaker
authors.affiliation: Institute of Parasitology, University of Bern, Bern, Switzerland; Department of Parasitology and Tropical Medicine, National Center of Infectious and Parasitic Diseases, Sofia, Bulgaria.
authors.forename: Irina
authors.initials: I
authors.lastname: Marinova
authors.affiliation: Institute of Parasitology, University of Zurich, Zurich, Switzerland.
authors.forename: Francesca
authors.initials: F
authors.lastname: Gori
authors.affiliation: Institute of Parasitology, University of Bern, Bern, Switzerland; University of Monastir, Faculty of Pharmacy, Department of Clinical Biology B, Laboratory of Medical and Molecular Parasitology-Mycology (LR12ES08), Monastir, Tunisia.
authors.forename: Amani
authors.initials: A
authors.lastname: Hizem
authors.affiliation: Institute of Parasitology, University of Bern, Bern, Switzerland.
authors.forename: Norbert
authors.initials: N
authors.lastname: Müller
authors.affiliation: Department of Infectious, Parasitic and Immunomediated Diseases, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, Italy.
authors.forename: Adriano
authors.initials: A
authors.lastname: Casulli
authors.affiliation: Pedro Kouri Institute, Autopista Novia del Mediodia, La Lisa, La Habana, Cuba.
authors.forename: Luis Enrique
authors.initials: LE
authors.lastname: Jerez Puebla
authors.affiliation: University of Monastir, Faculty of Pharmacy, Department of Clinical Biology B, Laboratory of Medical and Molecular Parasitology-Mycology (LR12ES08), Monastir, Tunisia.
authors.forename: Hamouda
authors.initials: H
authors.lastname: Babba
authors.affiliation: Institute of Parasitology, University of Bern, Bern, Switzerland. Electronic address: bruno.gottstein@vetsuisse.unibe.ch.
authors.forename: Bruno
authors.initials: B
authors.lastname: Gottstein
authors.affiliation: Institute of Parasitology, University of Bern, Bern, Switzerland.
authors.forename: Markus
authors.initials: M
authors.lastname: Spiliotis
mesh_terms: D000818:Animals; D001483:Base Sequence; D004269:DNA, Bacterial / Q000235:genetics; D004446:Echinococcus / Q000235:genetics* / Q000302:isolation & purification*; D060005:Genotyping Techniques; D006801:Humans; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods*; D020641:Polymorphism, Single Nucleotide / Q000235:genetics; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA / Q000379:methods*; D013621:Taenia / Q000235:genetics* / Q000302:isolation & purification*
chemical_list: D004269:DNA, Bacterial
keywords: 12S and 16S nuclear ribosomal RNA genes; Echinococcus spp.; PCR; Specific and genotypic identification; Taenia spp.