Korczak Bożena M, Stieber Regina, Emler Stefan, Burnens André P, Frey Joachim, Kuhnert Peter
Institute of Veterinary Bacteriology, University of Bern, CH-3012 Bern, Switzerland.
SmartGene, Zug, Switzerland.
Int J Syst Evol Microbiol. 2006 May;56(Pt 5):937-945. doi: 10.1099/ijs.0.64109-0.
The genus Campylobacter comprises 17 species, some of which are important animal and human pathogens. To gain more insight into the genetic relatedness of this genus and to improve the molecular tools available for diagnosis, a universal sequencing approach was established for the gene encoding the beta-subunit of RNA polymerase (rpoB) for the genus Campylobacter. A total of 59 strains, including the type strains of currently recognized species as well as field isolates, were investigated in the study. A primer set specific for Campylobacter species enabled straightforward amplification and sequencing of a 530 bp fragment of the rpoB gene. The 16S rRNA gene sequences of all of the strains were determined in parallel. A good congruence was obtained between 16S rRNA and rpoB gene sequence-based trees within the genus Campylobacter. The branching of the rpoB tree was similar to that of the 16S rRNA gene tree, even though a few discrepancies were observed for certain species. The resolution of the rpoB gene within the genus Campylobacter was generally much higher than that of the 16S rRNA gene sequence, resulting in a clear separation of most species and even some subspecies. The universally applicable amplification and sequencing approach for partial rpoB gene sequence determination provides a powerful tool for DNA sequence-based discrimination of Campylobacter species.
弯曲杆菌属包含17个种,其中一些是重要的动物和人类病原体。为了更深入了解该属的遗传相关性并改进现有的诊断分子工具,建立了一种通用的测序方法,用于对弯曲杆菌属RNA聚合酶β亚基(rpoB)编码基因进行测序。该研究共调查了59株菌株,包括目前已确认种的模式菌株以及野外分离株。一套针对弯曲杆菌属的引物能够直接扩增并测序rpoB基因的一个530 bp片段。同时测定了所有菌株的16S rRNA基因序列。在弯曲杆菌属内,基于16S rRNA和rpoB基因序列构建的树状图具有良好的一致性。尽管某些种存在一些差异,但rpoB基因树的分支与16S rRNA基因树相似。弯曲杆菌属内rpoB基因的分辨率通常远高于16S rRNA基因序列,从而能够清晰区分大多数种甚至一些亚种。用于部分rpoB基因序列测定的通用扩增和测序方法为基于DNA序列鉴别弯曲杆菌属物种提供了一个强大的工具。