Identification of mycobacterial species by comparative sequence analysis of the RNA polymerase gene (rpoB)
Identification of mycobacterial species by comparative sequence analysis of the RNA polymerase gene (rpoB)
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DOI：
10.1128/jcm.37.6.1714-1720.1999
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发表时间：
1999-06-01
影响因子：
9.4
通讯作者：
Kook, YH
中科院分区：
文献类型：
--
作者：
Kim, BJ;Lee, SH;Kook, YH
为了区分和鉴定分枝杆菌物种，研究了编码RNA聚合酶β亚基的rpoB基因。用PCR方法从44株分枝杆菌参考株和107株临床分离株中扩增出rpoB基因。直接测定核苷酸序列（306 bp），并通过使用MegAlign软件包（DNASTAR）中的多重比对算法和MEGA程序进行比对。采用邻接法构建系统发育树。rpoB DNA的比较序列分析为分枝杆菌属内的物种区分提供了依据。在系统发育树中，缓慢生长和快速生长的分枝杆菌群被清楚地分开，每个分枝杆菌种被区分为不同的实体，致病性堪萨斯分枝杆菌很容易与非致病性M. gastri;这种分化不能通过使用16 S rRNA基因（rDNA）序列来实现。通过将相同物种的菌株之间具有低水平序列差异的物种特异性聚类，可以容易地鉴定所有临床分离株。这些结果表明，扩增的rpoB DNA的比较序列分析可以有效地用于与传统的培养方法平行，并作为165 rDNA基因分析的补充，以确定分枝杆菌的临床分离株。此外，在M.结核病、利福平耐药性可同时测定。
For the differentiation and identification of mycobacterial species, the rpoB gene, encoding the beta subunit of RNA polymerase, was investigated. rpoB DNAs (342 bp) were amplified from 44 reference strains of mycobacteria and clinical isolates (107 strains) by PCR. The nucleotide sequences were directly determined (306 bp) and aligned by using the multiple alignment algorithm in the MegAlign package (DNASTAR) and the MEGA program. A phylogenetic tree was constructed by the neighbor-joining method. Comparative sequence analysis of rpoB DNAs provided the basis for species differentiation within the genus Mycobacterium. Slowly and rapidly growing groups of mycobacteria were clearly separated, and each mycobacterial species was differentiated as a distinct entity in the phylogenetic tree, Pathogenic Mycobacterium kansasii was easily differentiated from nonpathogenic M. gastri; this differentiation cannot be achieved by using 16S rRNA gene (rDNA) sequences. By being grouped into species-specific clusters with low-level sequence divergence among strains of the same species, all of the clinical isolates could be easily identified. These results suggest that comparative sequence analysis of amplified rpoB DNAs can be used efficiently to identify clinical isolates of mycobacteria in parallel with traditional culture methods and as a supplement to 165 rDNA gene analysis. Furthermore, in the case of M. tuberculosis, rifampin resistance can be simultaneously determined.