Minárik Gabriel, Bazsalovicsová Eva, Zvijáková Ludmila, Stefka Jan, Pálková Lenka, Králová-Hromadová Ivica
Department of Molecular Biology, Faculty of Natural Sciences, Comenius University, Mlynská dolina B-2, 84215 Bratislava, Slovakia; Institute of Molecular Biomedicine, Faculty of Medicine, Comenius University, Sasinkova 4, 81108 Bratislava, Slovakia; Geneton Ltd., Ilkovičova 3, 84104 Bratislava, Slovakia.
Institute of Parasitology, Slovak Academy of Sciences, Hlinkova 3, 04001 Košice, Slovakia.
Mol Biochem Parasitol. 2014 Jun;195(1):30-3. doi: 10.1016/j.molbiopara.2014.06.003. Epub 2014 Jun 21.
The microsatellite markers were designed for the giant liver fluke, Fascioloides magna, veterinary important liver parasite of free-living and domestic ruminants. Due to its geographic distribution (five enzootic regions across USA and Canada, three permanent European foci) and invasive character, F. magna is an interesting model for population genetics. Out of 667 amplicon candidates generated after NGS, 118 provided the best resolution and were tested with PCR analysis. In total, 56 yielded PCR products of expected size and in 36 of them the declared repetitive motif was identified by Sanger sequencing. After fragment analysis, 12 loci were proved to be polymorphic in individuals from one tested European and four North American populations. These loci were selected for setup of multiplex STR assays and utilized in genotyping of larger sample cohort. The outputs of statistical analyses indicate further global application of 11 conclusive loci in population genetics of the parasite.
微卫星标记是针对巨片形吸虫（Fascioloides magna）设计的，它是自由生活和家养反刍动物重要的肝脏寄生虫。由于其地理分布（美国和加拿大的五个动物疫病流行区，欧洲的三个永久性疫源地）以及入侵特性，巨片形吸虫是群体遗传学的一个有趣模型。在二代测序后产生的667个扩增子候选物中，118个具有最佳分辨率，并通过PCR分析进行了测试。总共56个产生了预期大小的PCR产物，其中36个通过桑格测序鉴定出了宣称的重复基序。经过片段分析，在一个测试的欧洲种群和四个北美种群的个体中，有12个位点被证明具有多态性。这些位点被选择用于建立多重STR分析，并用于更大样本队列的基因分型。统计分析结果表明，11个决定性位点在该寄生虫的群体遗传学中具有进一步的全球应用价值。
