A dual PCR-based sequencing approach for the identification and discrimination of Echinococcus and Taenia taxa

Research paper by Ghalia Boubaker, Irina Marinova, Francesca Gori, Amani Hizem, Norbert Müller, Adriano Casulli, Luis Enrique Jerez Puebla, Hamouda Babba, Bruno Gottstein, Markus Spiliotis

Indexed on: 28 May '16Published on: 27 May '16Published in: MCP


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 E. granulosus sensu stricto to be distinguished, and 25 SNPs that allowed differentiation within E. 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.

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