基于16S rRNA和rpoB基因的分子系统发育分析在铜绿假单胞菌鉴定中的应用

Molecular Phylogeny Analysis Based on 16S rRNA and rpoB Sequences for Identification of Clinical Isolates of Pseudomonas aeruginosa

为对比16S rRNA和rpo B基因分子系统发育分析与传统表型分类法对铜绿假单胞菌的鉴定,评估16S rRNA和rpo B基因序列分析在铜绿假单胞菌鉴定中的应用,用表型分类方法对临床自动微生物鉴定系统鉴定为铜绿假单胞菌的23株分离株进行再鉴定,PCR扩增23株分离株16S rRNA和rpo B基因片段,并测序进行系统发育分析。结果表明,表型再鉴定结果与自动微生物鉴定系统鉴定结果一致。基于两个基因的系统发育分析均显示分离株p22与不动杆菌属序列聚为一枝,其余22株分离株与铜绿假单胞菌序列聚为一枝。因此p22应鉴定为不动杆菌,16S rRNA和rpo B基因序列分析均能准确鉴定铜绿假单胞菌并能较好建立假单胞菌属内种间关系。

16S rRNA and rpo B gene sequence analysis to identify of Pseudomonas aeruginosa was compared with conventional phenotypic methods to evaluate the application of 16 S rRNA and rpo B sequence analysis to identify P.aeruginosa; 23 isolated strains that were re-characterized with phenotypic classification method that was identified as P.aeruginosa by clinical automatic microbial identification system.16 S rRNA and rpo B gene fragments were amplified with PCR and sequenced for molecular phylogenic analysis.The results showed that the re-characterized results were consistent with those characterized with automatic microbial identification system.Using Bayesian phylogenetic analyses based on 16 S rRNA and rpo B genes,22 isolates are characterized as P.aeruginosa,whereas one isolate,p22,was classified as Acinetobacter sp.p22 should be identified as Acinetobacter sp.16 S rRNA and rpo B genes can accurate identify P.aeruginosa and could fairly establish the interspecific relation within the genus of Pseudomonas.