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[从临床样本中分离出的屎肠球菌和粪肠球菌菌株的抗生素耐药性及毒力因子研究]

[Investigation of Antibiotic Resistance and Virulence Factors of Enterococcus faecium and Enterococcus faecalis Strains Isolated from Clinical Samples].

作者信息

Gök Şerife Merve, Türk Dağı Hatice, Kara Fatih, Arslan Uğur, Fındık Duygu

机构信息

Selcuk University Faculty of Medicine, Department of Medical Microbiology, Konya, Turkey.

Selcuk University Faculty of Medicine, Department of Public Health, Konya, Turkey.

出版信息

Mikrobiyol Bul. 2020 Jan;54(1):26-39. doi: 10.5578/mb.68810.

Abstract

Enterococci, which are commonly found in the environment, cause serious infections despite the absence of well-defined virulence factors and toxins. Knowing the virulence properties of enterococci is important to understand the complex pathogenic structures. In this study, we aimed to investigate the virulence factors (asa1, hyl, cylA, efa, ebp, ace, esp, gelE, sprE, fsrA, fsrB, fsrC genes, gelatinase activity, hemolysin, hydrogen peroxide and biofilm production) and antibiotic resistance of Enterococcus faecium and Enterococcus faecalis strains isolated from clinical specimens. A total of 110 enterococcus isolates which were accepted as infectious agents were included in the study. The polymerase chain reaction method was used to identify the isolates and to detect virulence genes. Characteristics of hemolysis, biofilm formation, hydrogen peroxide production and gelatinase activity were investigated by phenotypic methods. The antibiotic susceptibility test was performed with VITEK 2 automated system. E.faecalis ATCC 29212 standard strain was used as a quality control in all tests. Of the 110 enterococci isolates included in the study, 61 were identified as E.faecium and 49 as E.faecalis. The efa gene was the most frequently detected virulence gene (92.7%), followed by ace (83.6%), esp (66.4%), ebp (60.0%), cylA (50.9%), hyl (46.4%), asa1 (45.5%), gelE, sprE, fsrC (33.6%), fsrA (12.7%) and fsrB (11.8%). All genes except hyl were higher in E.faecalis isolates and the difference was statistically significant (p<0.05). Twenty-five (51%) E.faecalis and 1 (1.6%) E.faecium isolates had beta-hemolysis and the difference was statistically significant (p= 0.000). Seven (11.5%) E.faecium and 4 (8.2%) E.faecalis isolates formed biofilm, but the difference was not statistically significant (p> 0.05). Two (3.3%) E.faecium and 14 (28.6%) E.faecalis isolates exhibited gelatinase activity and the difference between the two species was statistically significant (p= 0.000). Hydrogen peroxide production was not detected in any of the isolates. The highest resistance rate was determined against ciprofloxacin (70.9%). The resistance to ampicillin was 69.1%, high level streptomycin 65.1%, high level gentamicin 39.4%, vancomycin and teicoplanin 4.5%, and linezolid 1.8%. In conclusion, our data indicated that virulence factors except hyl gene and biofilm production were higher in E.faecalis isolates but E.faecium isolates were more resistant to antibiotics. In order to prevent infection of such virulent or resistant isolates in the hospital setting, infection control measures must be followed. In vivo studies are needed for the better understanding of the virulence of enterococci.

摘要

肠球菌常见于环境中,尽管缺乏明确的毒力因子和毒素,但仍会引发严重感染。了解肠球菌的毒力特性对于理解其复杂的致病结构至关重要。在本研究中,我们旨在调查从临床标本中分离出的粪肠球菌和屎肠球菌菌株的毒力因子(asa1、hyl、cylA、efa、ebp、ace、esp、gelE、sprE、fsrA、fsrB、fsrC基因、明胶酶活性、溶血素、过氧化氢和生物膜形成)及抗生素耐药性。本研究共纳入110株被视为感染病原体的肠球菌分离株。采用聚合酶链反应方法鉴定分离株并检测毒力基因。通过表型方法研究溶血、生物膜形成、过氧化氢产生和明胶酶活性的特征。使用VITEK 2自动化系统进行抗生素敏感性试验。在所有试验中,粪肠球菌ATCC 29212标准菌株用作质量控制。在本研究纳入的110株肠球菌分离株中,61株被鉴定为屎肠球菌,49株为粪肠球菌。efa基因是最常检测到的毒力基因(92.7%),其次是ace(83.6%)、esp(66.4%)、ebp(60.0%)、cylA(50.9%)、hyl(46.4%)、asa1(45.5%)、gelE、sprE、fsrC(33.6%)、fsrA(12.7%)和fsrB(11.8%)。除hyl外,所有基因在粪肠球菌分离株中的检出率均更高,差异具有统计学意义(p<0.05)。25株(51%)粪肠球菌和1株(1.6%)屎肠球菌分离株具有β溶血,差异具有统计学意义(p = 0.000)。7株(11.5%)屎肠球菌和4株(8.2%)粪肠球菌分离株形成生物膜,但差异无统计学意义(p>0.05)。2株(3.3%)屎肠球菌和十四株(28.6%)粪肠球菌分离株表现出明胶酶活性,两种菌之间的差异具有统计学意义(p = 0.000)。在任何分离株中均未检测到过氧化氢产生。对环丙沙星的耐药率最高(70.9%)。对氨苄西林的耐药率为69.1%,对高水平链霉素为65.1%,对高水平庆大霉素为39.4%,对万古霉素和替考拉宁为4.5%,对利奈唑胺为1.8%。总之,我们的数据表明,除hyl基因外,粪肠球菌分离株中的毒力因子和生物膜形成更高,但屎肠球菌分离株对抗生素更耐药。为防止医院环境中此类有毒或耐药分离株的感染,必须遵循感染控制措施。需要进行体内研究以更好地了解肠球菌的毒力。

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