Lee S H, Kim B J, Kim J H, Park K H, Kim S J, Kook Y H
Department of Microbiology, College of Medicine, Konkuk University, Chungju, Chungchongbuk-Do 380-701, Korea.
J Clin Microbiol. 2000 Jul;38(7):2557-62. doi: 10.1128/JCM.38.7.2557-2562.2000.
We determined the nucleotide sequences (329 bp) of the rpoB DNAs from 22 reference strains of Borrelia. No insertions or deletions were observed. Deduced amino acid sequences of amplified rpoB DNA comprised 109 amino acid residues (N(450) to M(558) [Escherichia coli numbering]). All amino acid sequences were identical with the exception of those of Borrelia lusitaniae PotiB2 (T(461)-->A) and B. bissettii DN127 (I(498)-->V). Each species of B. burgdorferi sensu lato was differentiated as a distinct entity in the phylogenetic tree constructed by a neighbor-joining method. B. burgdorferi sensu lato could be distinguished from B. turicatae and B. hermsii, which are associated with relapsing fever. Seventeen Korean isolates could be identified by PCR-linked direct sequencing and restriction analysis of the rpoB DNA. These results suggest that rpoB DNA is useful for identification and characterization of Borrelia. In addition, we developed the rapid species identification method using the species-specific primer sets based on rpoB gene sequences.
我们测定了来自22株疏螺旋体参考菌株的rpoB DNA的核苷酸序列(329 bp)。未观察到插入或缺失。扩增的rpoB DNA推导的氨基酸序列包含109个氨基酸残基(N(450)至M(558) [大肠杆菌编号])。除了葡萄牙疏螺旋体PotiB2(T(461)-->A)和比氏疏螺旋体DN127(I(498)-->V)的氨基酸序列外,所有氨基酸序列均相同。在通过邻接法构建的系统发育树中,狭义伯氏疏螺旋体的每个物种都被区分为一个独特的实体。狭义伯氏疏螺旋体可与与回归热相关的图氏疏螺旋体和赫氏疏螺旋体区分开来。通过rpoB DNA的PCR连锁直接测序和限制性分析,可以鉴定出17株韩国分离株。这些结果表明,rpoB DNA可用于疏螺旋体的鉴定和特征描述。此外,我们基于rpoB基因序列开发了使用物种特异性引物对的快速物种鉴定方法。