Kuzucu Cigdem, Cizmeci Zeynep, Durmaz Riza, Durmaz Bengul, Ozerol Ibrahim Halil
Department of Clinical Microbiology, Inonu University, Malatya, Turkey.
Microb Drug Resist. 2005 Summer;11(2):159-64. doi: 10.1089/mdr.2005.11.159.
The gastrointestinal tract carriage of enterococci was searched in 150 hospitalized patients and 100 outpatients, and clonal relatedness of the isolates and their resistance to ampicillin, vancomycin, and high-level streptomycin and gentamicin were investigated. A stool sample or rectal swab collected from each patient was inoculated into appropriate media within an hour. Enterococcus species were identified by using conventional biochemical tests, API-20 Strep assay, and BBL crystal kit. Antibiotic susceptibility tests were performed using Kirby-Bauer disk diffusion method. A polymerase chain reaction (PCR) was used to detect vanA and vanB genes. Pulsed-field gel electrophoresis (PFGE) and arbitrarily primed-polymerase chain reaction (AP-PCR) methods were used for molecular typing of the strains. Enterococci were isolated from 90 (60%) of the specimens collected from 150 inpatients. Of these 90 isolates, 37 (41%) had high-level gentamicin resistance, 36 (40%) had high-level streptomycin resistance, and 50 (55.6%) had ampicillin resistance. Fecal colonization was found in 30% of the outpatients. Resistances to ampicillin, high-level streptomycin, and gentamicin were 13%, 10%, and 3%, in these patients' isolates, respectively. No vancomycin-resistant enterococci were detected by both agar diffusion and PCR assays in our study. Both typing procedures were applied on 78 Enterococcus strains isolated from inpatients. AP-PCR typing showed that 30 (50.8%) of the 59 E. faecium and 5 (50%) of the 10 E. faecalis strains were clonally related. These values were found to be 12 (20.3%) and two (20%) by PFGE, respectively. The typing procedures did not find any clustered strains in the six E. durans and three E. avium isolates. Neither PFGE nor AP-PCR result was significantly different among the sensitive and resistant strains. Our results indicate that the high prevalence of colonization with ampicillin and highlevel aminoglycoside-resistant enterococci is an important problem in our medical center. The high clonal diversity among the isolates indicates limited spread of antibiotic-resistant strains between patients.
在150名住院患者和100名门诊患者中对肠道球菌的胃肠道携带情况进行了调查,并研究了分离株的克隆相关性及其对氨苄西林、万古霉素、高水平链霉素和庆大霉素的耐药性。从每位患者采集的粪便样本或直肠拭子在1小时内接种到合适的培养基中。使用常规生化试验、API-20 Strep检测法和BBL晶体试剂盒鉴定肠球菌种类。采用 Kirby-Bauer 纸片扩散法进行药敏试验。使用聚合酶链反应(PCR)检测vanA和vanB基因。采用脉冲场凝胶电泳(PFGE)和任意引物聚合酶链反应(AP-PCR)方法对菌株进行分子分型。从150名住院患者采集的90份(60%)标本中分离出肠球菌。在这90株分离株中,37株(41%)对高水平庆大霉素耐药,36株(40%)对高水平链霉素耐药,50株(55.6%)对氨苄西林耐药。在30%的门诊患者中发现粪便定植。在这些患者的分离株中,对氨苄西林、高水平链霉素和庆大霉素的耐药率分别为13%、10%和3%。在我们的研究中,琼脂扩散法和PCR检测均未检测到耐万古霉素肠球菌。两种分型方法均应用于从住院患者中分离出的78株肠球菌菌株。AP-PCR分型显示,59株粪肠球菌中的30株(50.8%)和10株屎肠球菌中的5株(50%)具有克隆相关性。通过PFGE分别发现这些值为12株(20.3%)和2株(20%)。分型方法在6株耐久肠球菌和3株鸟肠球菌分离株中未发现任何聚集菌株。敏感菌株和耐药菌株之间的PFGE和AP-PCR结果均无显著差异。我们的结果表明,氨苄西林和高水平氨基糖苷类耐药肠球菌定植的高流行率是我们医疗中心的一个重要问题。分离株之间的高克隆多样性表明耐药菌株在患者之间的传播有限。