Nikmanesh Bahram, Mirhendi Hossein, Mahmoudi Shahram, Rokni Mohammad Bagher
Department of Medical Laboratory Science, School of Allied Medical Sciences, Tehran University of Medical Sciences, Tehran, Iran.
Department of Medical Parasitology & Mycology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran.
Exp Parasitol. 2017 Dec;183:50-55. doi: 10.1016/j.exppara.2017.10.002. Epub 2017 Oct 9.
Echinococcus granulosus is now considered a complex consisting of at least four species and ten genotypes. Different molecular targets have been described for molecular characterization of E. granulosus; however, in almost all studies only one or two of the targets have been used, and only limited data is available on the utilization of multiple loci. Therefore, we investigated the genetic diversity among 64 strains isolated from 138 cyst specimens of human and animal isolates, using a set of nuclear and mitochondrial genes; i.e., cytochrome c oxidase subunit 1 (cox1), NADH dehydrogenase subunit 1 (nad1), ATPase subunit 6 (atp6), 12S rRNA (12S), and Actin II (act II). In comparison to the use of molecular reference targets (nad1 + cox1), using singular target (act II or 12S or atp6) yielded lower discriminatory power. Act II and 12S genes could accurately discriminate the G6 genotype, but they were not able to differentiate between G1 and G3 genotypes. As the G1 and G3 genotypes belong to the E. granulosus sensu stricto, low intra-species variation was observed for act II and 12S. The atp6 gene could identify the G3 genotype but could not differentiate G6 and G1 genotypes. Using concatenated sequence of five genes (cox1 + nad1 + atp6 + 12S + act II), genotypes were identified accurately, and markedly higher resolution was obtained in comparison with the use of reference markers (nad1 + cox1) only. Application of multilocus sequence analysis (MLSA) to large-scale studies could provide valuable epidemiological data to make efficient control and management measures for cystic echinococcosis.
细粒棘球绦虫现在被认为是一个复合体，至少由四个物种和十个基因型组成。已经描述了用于细粒棘球绦虫分子特征分析的不同分子靶点；然而，在几乎所有研究中，仅使用了一两个靶点，关于多个位点利用的可用数据有限。因此，我们使用一组核基因和线粒体基因，即细胞色素c氧化酶亚基1（cox1）、NADH脱氢酶亚基1（nad1）、ATP酶亚基6（atp6）、12S核糖体RNA（12S）和肌动蛋白II（act II），研究了从138份人和动物分离的囊肿标本中分离出的64株菌株的遗传多样性。与使用分子参考靶点（nad1 + cox1）相比，使用单一靶点（act II或12S或atp6）产生的鉴别力较低。Act II和12S基因能够准确区分G6基因型，但无法区分G1和G3基因型。由于G1和G3基因型属于狭义的细粒棘球绦虫，act II和12S的种内变异较低。Atp6基因可以识别G3基因型，但无法区分G6和G1基因型。使用五个基因的串联序列（cox1 + nad1 + atp6 + 12S + act II）能够准确鉴定基因型，与仅使用参考标记（nad1 + cox1）相比，获得了明显更高的分辨率。将多位点序列分析（MLSA）应用于大规模研究可为制定有效的囊性棘球蚴病控制和管理措施提供有价值的流行病学数据。
