Shamsaddini Saeedeh, Mohammadi Mohammad Ali, Mirbadie Seyed Reza, Rostami Sima, Dehghani Mansoureh, Sadeghi Balal, Harandi Majid Fasihi
Kerman University of Medical Sciences, Research Center for Hydatid Disease in Iran, Kerman, Iran.
Kerman University of Medical Sciences, Sirjan School of Medical Sciences, Sirjan, Iran.
Rev Inst Med Trop Sao Paulo. 2017 Sep 4;59:e66. doi: 10.1590/S1678-9946201759066.
Canine taeniids are among the major tapeworms with remarkable medical and economic significance. Reliable diagnosis and differentiation of dog taeniids using simple and sensitive tools are of paramount importance for establishing an efficient surveillance system. Microsatellites as abundant unique tandem repeats of short DNA motifs are useful genetic markers for molecular epidemiological studies. The purpose of the present study was to find a primer pair for rapid differentiation of major tapeworms of dogs, Taenia hydatigena, T. multiceps, T. ovis and Echinococcus granulosus, by screening existing nucleotide data. All the mitochondrial genome records as well as non-coding ITS1 sequences of Taeniidae species were downloaded from Nucleotide database from NCBI. For prediction and analysis of potential loci of STR/SSR in ITS1 as well as mitochondrial regions, we used ChloroMitoSSRDB 2.0 and GMATo v1.2. software. Different tapeworm species were categorized according to different motif sequences and type and size of each microsatellite locus. Three primer sets were designed and tested for differentiating taeniid species and evaluated in a conventional PCR system. Four taeniid species were successfully differentiated using a primer pair in a simple conventional PCR system. We predicted 2-19 and 1-4 microsatellite loci in ITS1 and mitochondrial genome, respectively. In ITS1, 41 Di and 21 Tri motifs were found in the taeniids while the majority of the motifs in the mitochondrial genome were Tetra (89) and Tri (70). It is documented that the number and diversity of microsatellite loci is higher in nuclear ITS1 region than mostly coding mitochondrial genome.
犬绦虫是具有重要医学和经济意义的主要绦虫之一。使用简单且灵敏的工具对犬绦虫进行可靠的诊断和鉴别对于建立高效的监测系统至关重要。微卫星作为短DNA基序丰富的独特串联重复序列，是分子流行病学研究中有用的遗传标记。本研究的目的是通过筛选现有核苷酸数据，找到一对引物，用于快速鉴别犬的主要绦虫，即泡状带绦虫、多头带绦虫、羊带绦虫和细粒棘球绦虫。所有带科物种的线粒体基因组记录以及非编码ITS1序列均从NCBI的核苷酸数据库下载。为了预测和分析ITS1以及线粒体区域中STR/SSR的潜在位点，我们使用了ChloroMitoSSRDB 2.0和GMATo v1.2软件。根据不同的基序序列以及每个微卫星位点的类型和大小对不同的绦虫物种进行分类。设计并测试了三套引物用于鉴别绦虫物种，并在常规PCR系统中进行评估。在一个简单的常规PCR系统中，使用一对引物成功鉴别出四种绦虫物种。我们分别在ITS1和线粒体基因组中预测到2 - 19个和1 - 4个微卫星位点。在ITS1中，绦虫中发现了41个二核苷酸和21个三核苷酸基序，而线粒体基因组中的大多数基序是四核苷酸（89个）和三核苷酸（70个）。据记载，核ITS1区域中微卫星位点的数量和多样性高于大多数编码线粒体基因组。
