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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
引用本文:何艳霞,向诗非,李浇,何金蕾,张俊荣,袁冬梅,秦翰霄,陈达丽,陈建平.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[J].微生物学杂志,2016(4):27-35.
作者姓名:何艳霞  向诗非  李浇  何金蕾  张俊荣  袁冬梅  秦翰霄  陈达丽  陈建平
作者单位:1.四川大学 华西基础医学与法医学院寄生虫学教研室,四川 成都 610041;2.湖北民族学院 医学院寄生虫学教研室,湖北 恩施 445000,湖北民族学院附属民大医院 风湿免疫科,湖北 恩施 445000,四川大学 华西基础医学与法医学院寄生虫学教研室,四川 成都 610041,四川大学 华西基础医学与法医学院寄生虫学教研室,四川 成都 610041,四川大学 华西基础医学与法医学院寄生虫学教研室,四川 成都 610041,四川大学 华西基础医学与法医学院寄生虫学教研室,四川 成都 610041,四川大学 华西基础医学与法医学院寄生虫学教研室,四川 成都 610041,四川大学 华西基础医学与法医学院寄生虫学教研室,四川 成都 610041,四川大学 华西基础医学与法医学院寄生虫学教研室,四川 成都 610041
基金项目:国家自然科学基金项目(31572240, 81171607, J1103604);四川大学与泸州市合作基金(2013CDLZ-S16)
摘    要: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。

关 键 词:铜绿假单胞菌  16S  rRNA  rpoB

Molecular Phylogeny Analysis Based on 16S rRNA and rpoB Sequences forIdentification of Clinical Isolates of Pseudomonas aeruginosa
HE Yan-xi,XIANG Shi-fei,LI Jiao,HE Jin-lei,ZHANG Jun-rong,YUAN Dong-mei,QIN Han-xiao,CHEN Da-li and CHEN Jian-ping.Molecular Phylogeny Analysis Based on 16S rRNA and rpoB Sequences forIdentification of Clinical Isolates of Pseudomonas aeruginosa[J].Journal of Microbiology,2016(4):27-35.
Authors:HE Yan-xi  XIANG Shi-fei  LI Jiao  HE Jin-lei  ZHANG Jun-rong  YUAN Dong-mei  QIN Han-xiao  CHEN Da-li and CHEN Jian-ping
Institution:1. Teach. & Res. Div.. of Parasitol., W. China Schl. of Basic & Forensic Med., Sichuan Uni., Chengdu 610041;2. Teach. & Res.Div. of Parasitol.,Coll of Med.,Hubei coll for Minornty Nationa lities,2. Teach. & Res.,Affil. Minda Hosp., Hubei Uni. for National''s, Enshi 445000,Teach. & Res. Div.. of Parasitol., W. China Schl. of Basic & Forensic Med., Sichuan Uni., Chengdu 610041,Teach. & Res. Div.. of Parasitol., W. China Schl. of Basic & Forensic Med., Sichuan Uni., Chengdu 610041,Teach. & Res. Div.. of Parasitol., W. China Schl. of Basic & Forensic Med., Sichuan Uni., Chengdu 610041,Teach. & Res. Div.. of Parasitol., W. China Schl. of Basic & Forensic Med., Sichuan Uni., Chengdu 610041,Teach. & Res. Div.. of Parasitol., W. China Schl. of Basic & Forensic Med., Sichuan Uni., Chengdu 610041 and Teach. & Res. Div.. of Parasitol., W. China Schl. of Basic & Forensic Med., Sichuan Uni., Chengdu 610041
Abstract:16S rRNA and rpoB gene sequence analysis to identify of Pseudomonas aeruginosa was compared with conventional phenotypic methods to evaluate the application of 16S rRNA and rpoB 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. 16S rRNA and rpoB 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 16S rRNA and rpoB genes, 22 isolates are characterized as P. aeruginosa, whereas one isolate, p22, was classified as Acinetobacter sp.p22 should be identified as Acinetobacter sp.16S rRNA and rpoB genes can accurate identify P. aeruginosa and could fairly establish the interspecific relation within the genus of Pseudomonas.
Keywords:Pseudomonas aeruginosa  16S rRNA  rpoB
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