Mukhacheva Tatyana A, Salikhova Irina I, Kovalev Sergey Y
Laboratory of Molecular Genetics, Department of Biology, Ural Federal University, Lenin Avenue 51, Yekaterinburg 620000, Russia.
Laboratory of Molecular Genetics, Department of Biology, Ural Federal University, Lenin Avenue 51, Yekaterinburg 620000, Russia.
Infect Genet Evol. 2015 Apr;31:257-62. doi: 10.1016/j.meegid.2015.02.009. Epub 2015 Feb 16.
Borrelia miyamotoi, a member of the relapsing fever group borreliae, was first isolated in Japan and subsequently found in Ixodes ticks in North America, Europe and Russia. Currently, there are three types of B. miyamotoi: Asian or Siberian (transmitted mainly by Ixodes persulcatus), European (Ixodesricinus) and American (Ixodesscapularis and Ixodespacificus). Despite the great genetic distances between B. miyamotoi types, isolates within a type are characterised by an extremely low genetic variability. In particular, strains of B. miyamotoi of Asian type, isolated in Russia from the Baltic sea to the Far East, have been shown to be identical based on the analysis of several conventional genetic markers, such as 16S rRNA, flagellin, outer membrane protein p66 and glpQ genes. Thus, protein or rRNA - coding genes were shown not to be informative enough in studying genetic diversity of B. miyamotoi within a type. In the present paper, we have attempted to design a new multilocus technique based on eight non-coding intergenic spacers (3686bp in total) and have applied it to the analysis of intra-type genetic variability of В. miyamotoi detected in different regions of Russia and from two tick species, I. persulcatus and Ixodespavlovskyi. However, even though potentially the most variable loci were selected, no genetic variability between studied DNA samples was found, except for one nucleotide substitution in two of them. The sequences obtained were identical to those of the reference strain FR64b. Analysis of the data obtained with the GenBank sequences indicates a highly homogeneous genetic background of B. miyamotoi from the Baltic Sea to the Japanese Islands. In this paper, a hypothesis of clonal expansion of B. miyamotoi is discussed, as well as possible mechanisms for the rapid dissemination of one B. miyamotoi clone over large distances.
宫本疏螺旋体是回归热疏螺旋体属的成员,最初在日本分离得到,随后在北美、欧洲和俄罗斯的硬蜱中被发现。目前,宫本疏螺旋体有三种类型:亚洲型或西伯利亚型(主要由全沟硬蜱传播)、欧洲型(蓖麻硬蜱)和美洲型(肩突硬蜱和太平洋硬蜱)。尽管宫本疏螺旋体各类型之间存在很大的遗传距离,但同一类型内的分离株具有极低的遗传变异性。特别是,从俄罗斯波罗的海到远东地区分离得到的亚洲型宫本疏螺旋体菌株,通过对几种传统遗传标记(如16S rRNA、鞭毛蛋白、外膜蛋白p66和glpQ基因)的分析显示是相同的。因此,蛋白质或rRNA编码基因在研究宫本疏螺旋体同一类型内的遗传多样性方面信息不足。在本文中,我们尝试设计一种基于八个非编码基因间隔区(共3686bp)的新多位点技术,并将其应用于分析在俄罗斯不同地区以及从全沟硬蜱和帕氏硬蜱这两种蜱中检测到的宫本疏螺旋体的型内遗传变异性。然而,尽管选择了潜在变异性最大的位点,但在所研究的DNA样本之间未发现遗传变异性,只有其中两个样本有一个核苷酸替换。获得的序列与参考菌株FR64b的序列相同。对从GenBank序列获得的数据的分析表明,从波罗的海到日本群岛的宫本疏螺旋体具有高度一致的遗传背景。本文讨论了宫本疏螺旋体克隆扩增的假说以及一个宫本疏螺旋体克隆远距离快速传播的可能机制。