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首页> 外文期刊>Applied Microbiology >Development of a Multilocus Sequence Typing Tool for High-Resolution Genotyping of Enterocytozoon bieneusi
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Development of a Multilocus Sequence Typing Tool for High-Resolution Genotyping of Enterocytozoon bieneusi

机译：高分辨率的Enterocytozoon bieneusi基因分型的多基因座序列分型工具的开发。

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Thus far, genotyping of Enterocytozoon bieneusi has been based solely on DNA sequence analysis of the internal transcribed spacer (ITS) of the rRNA gene. Both host-adapted and zoonotic (human-pathogenic) genotypes of E. bieneusi have been identified. In this study, we searched for microsatellite and minisatellite sequences in the whole-genome sequence database of E. bieneusi isolate H348. Seven potential targets (MS1 to MS7) were identified. Testing of the seven targets by PCR using two human-pathogenic E. bieneusi genotypes (A and Peru10) led to the selection of four targets (MS1, MS3, MS4, and MS7). Further analysis of the four loci with an additional 24 specimens of both host-adapted and zoonotic E. bieneusi genotypes indicated that most host-adapted genotypes were not amplified by PCR targeting these loci. In contrast, 10 or 11 of the 13 specimens of the zoonotic genotypes were amplified by PCR at each locus. Altogether, 12, 8, 7, and 11 genotypes of were identified at MS1, MS3, MS4, and MS7, respectively. Phylogenetic analysis of the nucleotide sequences obtained produced a genetic relationship that was similar to the one at the ITS locus, with the formation of a large group of zoonotic genotypes that included most E. bieneusi genotypes in humans. Thus, a multilocus sequence typing tool was developed for high-resolution genotyping of E. bieneusi. Data obtained in the study should also have implications for understanding the taxonomy of Enterocytozoon spp., the public health significance of E. bieneusi in animals, and the sources of human E. bieneusi infections.
机译：到目前为止，Enterocytozoon bieneusi的基因分型仅基于rRNA基因内部转录间隔区（ITS）的DNA序列分析。已经鉴定了比氏大肠杆菌的宿主适应型和人畜共患（人类致病）基因型。在这项研究中，我们在比氏大肠杆菌分离株H348的全基因组序列数据库中搜索了微卫星和微卫星序列。确定了七个潜在目标（MS1至MS7）。使用两种人类致病性比氏大肠杆菌基因型（A和Peru10）通过PCR对七个靶标进行测试，导致选择了四个靶标（MS1，MS3，MS4和MS7）。进一步分析四个基因座，再加上其他24个适应宿主和人畜共患的比埃希氏大肠杆菌基因型的标本，表明大多数适应宿主的基因型没有通过靶向这些基因座的PCR进行扩增。相反，在每个位点通过PCR扩增了人畜共患病基因型的13个样本中的10个或11个。分别在MS1，MS3，MS4和MS7上分别鉴定了12、8、7和11个基因型。对获得的核苷酸序列进行系统进化分析，得出了与ITS基因座相似的遗传关系，形成了一大群人畜共患病基因型，其中包括人类中大多数的比氏大肠杆菌基因型。因此，开发了一种多位点序列分型工具，用于高分辨率比氏大肠杆菌的基因分型。从研究中获得的数据也应该对理解肠小肠虫的分类，对动物中的比埃希氏大肠杆菌的公共卫生意义以及人类比氏埃希氏菌感染的来源具有启示意义。

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