Çopur Çiçek Ayşegül, Sandallı Cemal, Budak Emine Esra, Yağmur Gülhan, Çizmeci Zeynep, Ak Sibel, Balcı Pervin Özlem, Şay Coşkun Safiye Umut, Ay Altıntop Yasemin, Fırat Mehmet, Sarı Fatma, Çalışkan Ahmet, Yıldız Nazan, Sancaktar Metin, Özgümüş Osman Birol
Recep Tayyip Erdogan University Faculty of Medicine, Department of Medical Microbiology, Rize, Turkey.
Mikrobiyol Bul. 2016 Apr;50(2):175-85.
Escherichia coli is the most common pathogen isolated from both nosocomial and community acquired urinary tract infections. Although there are many studies from different centers concerning the antibiotic susceptibility of E.coli isolates in Turkey, the studies are quite few about class 1 and class 2 integron cassettes in clinical E.coli isolates from urinary samples. The aim of the study was to investigate the antibiotic susceptibility and the carriage of integron gene cassettes in E.coli strains isolated from urinary samples. A total of 626 E.coli strains isolated from urine cultures in microbiology laboratories located at 10 provinces from different regions of Turkey (Denizli, Ankara, Kayseri, Niğde, Şanlıurfa, Kahramanmaras, Tokat, Malatya, Konya and Trabzon) between June 2011-June 2012 were included in the study. The identification and antibiotic susceptibility testing of the isolates were studied by conventional methods as well as Vitek® 2 Compact (bioMérieux, France) and BD Phoenix™ 100 (Becton Dickinson, USA) systems. The antibiotic susceptibilities of all the isolates were retested by Kirby-Bauer disk diffusion method according to CLSI recommendations in the main center of the study in order to achive the standardization. The presence of integrons was detected with polymerase chain reaction (PCR) method by using specific primers targeting class 1 (intI1) and class 2 (intI2) integrase gene regions. After integron amplification the samples were cloned and subjected to DNA sequencing. When the antibiotic susceptibility of the isolates were evaluated, the highest resistance was observed against most commonly used empirical antibiotics namely ampicillin and trimethoprim-sulfamethoxazole (SXT) with the mean rate of 58.6% (range: 43.8%-73.2%) and 41.2% (range: 35.4%-45.8%), respectively. The most effective antibiotics detected against the isolates were imipenem and amikacin with the lowest resistance rates of 0.2% (range: 0%-1.1%) and 0.6% (range: 0%-3.2%), respectively. The frequency of positive IntI1 gene and class 1 integron gene cassettes were found as 25.8% (162/626) and 16.6% (104/626), respectively, whereas the frequency of positive intI2 gene II and class 2 integron gene cassettes were 5.1% (32/626) and 3% (19/626), respectively. The lowest intI1 gene frequency was detected in the isolates from Kayseri (16.6%) and the highest in the isolates from Kahramanmaraş (35.4%) provinces. While there was no intI2 gene in the isolates from Denizli and Kayseri, the highest frequency was 12.1% in the isolates from Şanlıurfa province. dfrA1 gene, the most frequent gene among integron gene cassettes was positive in 31 class 1 integron gene cassette alone, and positive with aadA1 gene in 18 class 1 integron gene cassettes. dfrA1 gene was positive with aadA1a just in one isolate. dfrA17 allele was positive in one isolate alone, in 28 isolates with aadA1, and in 15 isolates with aadA5. aadA1 gene was detected in four isolates. dfrA17-sat-aadA5 co-existence was detected among class 2 integron gene cassette in isolates from six provinces. dfrA1-sat-aadA1 was detected in one isolate from Ankara province and dfrA1 was detected in one isolate in Niğde province only. As a result, dfrA1 and aadA1 genes are the most common types of genes among class 1 and class 2 integron gene cassettes in E.coli isolated from urine cultures. It was concluded that high resistance against streptomycin (31.2%) and SXT (41.2%) supported the dissemination of integron-mediated genes dfr, sul1 and aad in the isolates.
大肠埃希菌是从医院获得性和社区获得性尿路感染中分离出的最常见病原体。尽管土耳其不同中心有许多关于大肠埃希菌分离株抗生素敏感性的研究,但关于尿液样本临床大肠埃希菌分离株中1类和2类整合子盒式结构的研究却相当少。本研究的目的是调查从尿液样本中分离的大肠埃希菌菌株的抗生素敏感性和整合子基因盒式结构的携带情况。2011年6月至2012年6月期间,从土耳其不同地区(代尼兹利、安卡拉、开塞利、尼德、尚勒乌尔法、卡拉曼马拉什、托卡特、马拉蒂亚、科尼亚和特拉布宗)10个省份的微生物实验室尿液培养物中分离出的626株大肠埃希菌菌株纳入了本研究。采用常规方法以及Vitek® 2 Compact(法国生物梅里埃公司)和BD Phoenix™ 100(美国贝克顿·迪金森公司)系统对分离株进行鉴定和抗生素敏感性测试。为实现标准化,在研究的主要中心根据CLSI建议通过 Kirby-Bauer纸片扩散法对所有分离株的抗生素敏感性进行重新检测。使用针对1类(intI1)和2类(intI2)整合酶基因区域的特异性引物,通过聚合酶链反应(PCR)方法检测整合子的存在。整合子扩增后,对样本进行克隆并进行DNA测序。当评估分离株的抗生素敏感性时,观察到对最常用的经验性抗生素氨苄西林和甲氧苄啶-磺胺甲恶唑(SXT)的耐药率最高,平均耐药率分别为58.6%(范围:43.8%-73.2%)和41.2%(范围:35.4%-45.8%)。检测到对分离株最有效的抗生素是亚胺培南和阿米卡星,最低耐药率分别为0.2%(范围:0%-1.1%)和0.6%(范围:0%-3.2%)。发现IntI1基因和1类整合子基因盒式结构的阳性频率分别为25.8%(162/626)和16.6%(104/626),而intI2基因II和2类整合子基因盒式结构的阳性频率分别为5.1%(32/626)和3%(19/626)。在开塞利分离株中检测到最低的intI1基因频率(16.6%),在卡拉曼马拉什省分离株中检测到最高频率(35.4%)。在代尼兹利和开塞利的分离株中没有intI2基因,而在尚勒乌尔法省分离株中最高频率为12.1%。整合子基因盒式结构中最常见的基因dfrA1基因,仅在31个1类整合子基因盒式结构中呈阳性,在18个1类整合子基因盒式结构中与aadA1基因同时呈阳性。dfrA1基因仅在一株分离株中与aadA1a同时呈阳性。dfrA17等位基因仅在一株分离株中呈阳性,在28株与aadA1同时存在的分离株中呈阳性,在15株与aadA5同时存在的分离株中呈阳性。在4株分离株中检测到aadA1基因。在6个省份分离株的2类整合子基因盒式结构中检测到dfrA17-sat-aadA5共存。在安卡拉省的一株分离株中检测到dfrA1-sat-aadA1,仅在尼德省的一株分离株中检测到dfrA1。结果,dfrA1和aadA1基因是从尿液培养物中分离的大肠埃希菌1类和2类整合子基因盒式结构中最常见的基因类型。得出结论,对链霉素(31.2%)和SXT(41.2%)的高耐药性支持了整合子介导的基因dfr、sul1和aad在分离株中的传播。