使用基于重复元件的聚合酶链反应分析法对患有囊性纤维化的成年和儿科患者所携带的铜绿假单胞菌分离株进行快速基因分型。
Rapid genotyping of Pseudomonas aeruginosa isolates harboured by adult and paediatric patients with cystic fibrosis using repetitive-element-based PCR assays.
作者信息
Syrmis Melanie W, O'Carroll Mark R, Sloots Theo P, Coulter Chris, Wainwright Claire E, Bell Scott C, Nissen Michael D
机构信息
Clinical Virology and Molecular Microbiology Research Unit, Sir Albert Sakzewksi Virus Research Centre, Royal Children's Hospital and Clinical Medical Virology Centre, University of Queensland, Herston, Queensland, Australia 4029 2,6Department of Paediatrics and Child Health2 and Department of Medicine6, University of Queensland, Brisbane, Queensland, Australia 3Adult Cystic Fibrosis Unit, The Prince Charles Hospital, Brisbane, Queensland, Australia 4Department of Respiratory Medicine, Royal Children's Hospital, Brisbane, Queensland, Australia 5Department of Microbiology, Queensland Health Pathology Service, The Prince Charles Hospital Campus, Brisbane, Queensland, Australia.
出版信息
J Med Microbiol. 2004 Nov;53(Pt 11):1089-1096. doi: 10.1099/jmm.0.45611-0.
In this study, the suitability of two repetitive-element-based PCR (rep-PCR) assays, enterobacterial repetitive intergenic consensus (ERIC)-PCR and BOX-PCR, to rapidly characterize Pseudomonas aeruginosa strains isolated from patients with cystic fibrosis (CF) was examined. ERIC-PCR utilizes paired sequence-specific primers and BOX-PCR a single primer that target highly conserved repetitive elements in the P. aeruginosa genome. Using these rep-PCR assays, 163 P. aeruginosa isolates cultured from sputa collected from 50 patients attending an adult CF clinic and 50 children attending a paediatric CF clinic were typed. The results of the rep-PCR assays were compared to the results of PFGE. All three assays revealed the presence of six major clonal groups shared by multiple patients attending either of the CF clinics, with the dominant clonal group infecting 38 % of all patients. This dominant clonal group was not related to the dominant clonal group detected in Sydney or Melbourne (pulsotype 1), nor was it related to the dominant groups detected in the UK. In all, PFGE and rep-PCR identified 58 distinct clonal groups, with only three of these shared between the two clinics. The results of this study showed that both ERIC-PCR and BOX-PCR are rapid, highly discriminatory and reproducible assays that proved to be powerful surveillance screening tools for the typing of clinical P. aeruginosa isolates recovered from patients with CF.
在本研究中,检测了两种基于重复元件的聚合酶链反应(rep-PCR)分析方法,即肠道细菌重复基因间共有序列(ERIC)-PCR和BOX-PCR,用于快速鉴定从囊性纤维化(CF)患者中分离出的铜绿假单胞菌菌株的适用性。ERIC-PCR使用一对序列特异性引物,而BOX-PCR使用单个引物,它们靶向铜绿假单胞菌基因组中高度保守的重复元件。使用这些rep-PCR分析方法,对从一家成人CF诊所的50名患者和一家儿科CF诊所的50名儿童痰液中培养出的163株铜绿假单胞菌分离株进行了分型。将rep-PCR分析的结果与脉冲场凝胶电泳(PFGE)的结果进行了比较。所有三种分析方法均显示,两家CF诊所的多名患者中存在六个主要克隆群,其中占主导地位的克隆群感染了所有患者的38%。这个占主导地位的克隆群与在悉尼或墨尔本检测到的主导克隆群(脉冲型1)无关,也与在英国检测到的主导克隆群无关。总体而言,PFGE和rep-PCR共鉴定出58个不同的克隆群,其中只有三个克隆群在两家诊所中共有。本研究结果表明,ERIC-PCR和BOX-PCR都是快速、高度鉴别且可重复的分析方法,被证明是用于对从CF患者中分离出的临床铜绿假单胞菌分离株进行分型的强大监测筛选工具。