Ferjani Sana, Saidani Mabrouka, Ennigrou Samir, Hsairi Mohamed, Slim Amine Faouzi, Ben Boubaker Ilhem Boutiba
Laboratory of Research, LR99ES09, Resistance aux Antimicrobiens, Faculty of Medicine of Tunis, University El manar of Tunis, 1007, Tunis, Tunisia,
Folia Microbiol (Praha). 2014 May;59(3):257-62. doi: 10.1007/s12223-013-0292-0. Epub 2013 Nov 21.
A collection of 201 Escherichia coli strains isolated from urine of patients in a Tunisian hospital between January 2006 and July 2008 was studied. Microbial identification was done by conventional methods, and antibiotic susceptibility with disk diffusion method was performed according to the Clinical Laboratory and Standards Institute guidelines. Detection of extended-spectrum beta-lactamase (ESBL) was performed by double-disk synergy test (DDST) and identification was done by PCR and sequencing. ESBL-producing isolates were subjected to molecular typing by random amplified polymorphic DNA (RAPD) and ST131 detection by PCR. Four phylogenetic groups (A, B1, B2 and D), 18 virulence genes and CTX-M group were individualized using PCR. Statistical analysis was done by Pearson χ2 test and Mann-Whitney U test. The strains were recovered primarily from urology (28%), maternity (19%) and medicine (16%) wards. Antibiotic resistance rates were ampicilin (72.1%), nalidixic acid (41.8%), ciprofloxacin (38.8%), gentamicin (23.9%) and cefotaxime (17.4%). Thirty-one of cefotaxime-resistant isolates (n = 35) had a positive DDST and harboured bla CTX-M-15 gene. Twenty of them (64.5%) belonged to the ST131 clone and showed the same RAPD DNA profile. Ciprofloxacin- and cotrimoxazole-susceptible isolates were significantly associated with phylogenetic group B2, whereas isolates that were resistant to these molecules were associated with B1 and D phylogenetic groups, respectively. Virulence genes were significantly more frequent among ciprofloxacin- and cotrimoxazole-susceptible strains than those resistant to these antibiotics. However, CXT-M-15-producing isolates were associated with many virulence genes. Isolates concomitantly susceptible to the three antimicrobials agents (ciprofloxacin, cefotaxime and cotrimoxazole) were significantly associated with group B2 and high virulence score, whereas isolates with resistance patterns especially those including resistance to ciprofloxacin belonged predominantly to B1 phylogroup and haboured few virulence genes. The emergence of virulent and multidrug-resistant E. coli is a concerning development that deserves close attention in our institution.
对2006年1月至2008年7月期间从突尼斯一家医院患者尿液中分离出的201株大肠杆菌菌株进行了研究。通过传统方法进行微生物鉴定,并根据临床实验室标准协会指南采用纸片扩散法进行抗生素敏感性检测。通过双纸片协同试验(DDST)检测超广谱β-内酰胺酶(ESBL),并通过PCR和测序进行鉴定。对产ESBL的分离株进行随机扩增多态性DNA(RAPD)分子分型,并通过PCR检测ST131。使用PCR区分四个系统发育组(A、B1、B2和D)、18个毒力基因和CTX-M组。通过Pearson χ2检验和Mann-Whitney U检验进行统计分析。这些菌株主要从泌尿外科(28%)、产科(19%)和内科(16%)病房分离得到。抗生素耐药率分别为氨苄西林(72.1%)、萘啶酸(41.8%)、环丙沙星(38.8%)、庆大霉素(23.9%)和头孢噻肟(17.4%)。35株对头孢噻肟耐药的分离株中有31株DDST呈阳性,并携带bla CTX-M-15基因。其中20株(64.5%)属于ST131克隆,且显示相同的RAPD DNA图谱。对环丙沙星和复方新诺明敏感的分离株与系统发育组B2显著相关,而对这些分子耐药的分离株分别与B1和D系统发育组相关。在对环丙沙星和复方新诺明敏感的菌株中,毒力基因的出现频率显著高于对这些抗生素耐药的菌株。然而,产CXT-M-15的分离株与许多毒力基因相关。对三种抗菌药物(环丙沙星、头孢噻肟和复方新诺明)均敏感的分离株与B2组和高毒力评分显著相关,而具有耐药模式的分离株,尤其是那些对环丙沙星耐药的分离株主要属于B1系统发育组,且毒力基因较少。毒性和多重耐药大肠杆菌的出现是一个令人担忧的发展情况,值得我们机构密切关注。