Boubaker Ghalia, Marinova Irina, Gori Francesca, Hizem Amani, Müller Norbert, Casulli Adriano, Jerez Puebla Luis Enrique, Babba Hamouda, Gottstein Bruno, Spiliotis Markus
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.
Institute of Parasitology, University of Bern, Bern, Switzerland; Department of Parasitology and Tropical Medicine, National Center of Infectious and Parasitic Diseases, Sofia, Bulgaria.
Mol Cell Probes. 2016 Aug;30(4):211-217. doi: 10.1016/j.mcp.2016.05.004. Epub 2016 May 27.
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.
用于棘球绦虫物种和/或基因型的遗传鉴定及区分的可靠且快速的分子工具，对于研究时空传播动态至关重要。在此，我们描述了一种针对线粒体核糖体RNA（rRNA）基因小亚基（rrnS）和大亚基（rrnL）区域的新型双重PCR方法，该方法能够：（i）特异性鉴定棘球绦虫的物种和基因型（rrnS + L-PCR），以及（ii）鉴定一系列带绦虫，包括不同种类的棘球绦虫、带绦虫及其他一些绦虫（17种双叶目蠕虫）。这种双重PCR方法具有高度敏感性，对棘球绦虫基因组DNA的分析检测限为1 pg。利用来自两个基因标记（1225 bp）的串联序列数据，我们鉴定出五个独特且具有地理信息的单核苷酸多态性（SNP），可区分狭义细粒棘球绦虫的基因型（G1和G3），以及25个可在加拿大棘球绦虫（G6/7/8/10）内进行区分的SNP。总之我们认为，这种基于双重PCR的测序方法可用于棘球绦虫和其他带绦虫的分子流行病学研究。
