Kang M, Xie Y, He C, Chen Z X, Guo L, Yang Q, Liu J Y, Du Y, Ou Q S, Wang L L
Department of Laboratory Medicine, West China Hospital, Sichuan University, Chengdu, 610041, China.
Eur J Clin Microbiol Infect Dis. 2014 Jun;33(6):933-9. doi: 10.1007/s10096-013-2029-z. Epub 2014 Jan 24.
The aim of this study was to characterize vancomycin-resistant Enterococcus faecium (VREfm) isolates phenotypically and molecularly, and investigate associations between the virulence factors enterococcal surface protein (esp), hyaluronidase (hyl), and collagen adhesin (acm) and colonization/infection. A total of 126 E. faecium [66 VREfm and 60 vancomycin-susceptible (VSEfm)] were collected in West China Hospital. Nine E. faecium isolates (7 VREfm and 2 VSEfm) were selected at random for comparative study in a large region from China. Minimum inhibitory concentrations (MICs) were measured by Etest and agar dilution, vancomycin resistance genes (vanA, vanB, and vanC) and virulence genes (esp, acm, and hyl) were detected by polymerase chain reaction (PCR). Thirty-four VREfm underwent repetitive sequence-based PCR (rep-PCR) and multi-locus sequence typing (MLST). One linezolid-resistant isolate (MIC = 8 μg/ml) was found; none were tigecycline resistant. All 73 VREfm (28 infective strains and 45 intestinal colonizers) had the vanA gene and VanA phenotype. Positivity for esp, hyl, and acm in VREfm was 79.5, 46.6, and 86.3%, respectively, which was higher than in VSEfm (54.8, 27.4, and 56.5%, respectively). Among VSEfm, positivity for acm in isolates from pleural or cerebrospinal fluid (84.6%) was higher than that from blood (32.4%). There were 11 rep-PCR types (similarity >95%) and MLST revealed nine sequence types (STs) among the selected isolates. Most VREfm and all VSEfm belonged to clonal complex 17. A new ST was found, with allele sequence (15, 1, 38, 1, 1, 1, 1). In China, most VREfm seem to belong to the classical nosocomial CC17 clone, and many of them have acquired virulence genes, further strengthening a hospital-adapted type.
本研究旨在从表型和分子水平对耐万古霉素屎肠球菌(VREfm)分离株进行特征分析,并研究毒力因子肠球菌表面蛋白(esp)、透明质酸酶(hyl)和胶原黏附素(acm)与定植/感染之间的关联。在华西医院共收集了126株屎肠球菌[66株VREfm和60株万古霉素敏感(VSEfm)]。随机选择9株屎肠球菌分离株(7株VREfm和2株VSEfm)在中国的一个大区域进行比较研究。采用Etest和琼脂稀释法测定最低抑菌浓度(MIC),通过聚合酶链反应(PCR)检测万古霉素耐药基因(vanA、vanB和vanC)和毒力基因(esp、acm和hyl)。对34株VREfm进行基于重复序列的PCR(rep-PCR)和多位点序列分型(MLST)。发现1株耐利奈唑胺分离株(MIC = 8 μg/ml);无耐替加环素的分离株。所有73株VREfm(28株感染菌株和45株肠道定植菌)均具有vanA基因和VanA表型。VREfm中esp、hyl和acm的阳性率分别为79.5%、46.6%和86.3%,高于VSEfm(分别为54.8%、27.4%和56.5%)。在VSEfm中,来自胸腔或脑脊液的分离株中acm的阳性率(84.6%)高于血液来源的分离株(32.4%)。在所选分离株中,有11种rep-PCR类型(相似度>95%),MLST显示有9种序列类型(STs)。大多数VREfm和所有VSEfm属于克隆复合体17。发现了一个新的ST,其等位基因序列为(15, 1, 38, 1, 1, 1, 1)。在中国,大多数VREfm似乎属于经典的医院内CC17克隆,其中许多已获得毒力基因,进一步强化了一种适应医院环境的类型。