Eser Ozgen Köseoğlu, Altun Uludağ Hatice, Ergin Alper, Boral Barış, Sener Burçin, Hasçelik Gülşen
Hacettepe University Faculty of Medicine, Department of Medical Microbiology, Ankara, Turkey.
Mikrobiyol Bul. 2014 Jan;48(1):59-69.
The aim of this study was to investigate the presence of carbapenem resistance in Enterobacteriaeceae isolates recovered from invasive infections, in Hacettepe University Hospital, Ankara, Turkey, between 2005-2009, by phenotypic and genotypic methods. A total of 210 non-duplicated Escherichia coli (n= 153), Klebsiella pneumoniae (n= 47) and Klebsiella oxytoca (n= 10) isolates which were all determined to be extended-spectrum beta-lactamase (ESBL) positive with the BD Phoenix automated identification and antibiotic susceptibility system (Sparks, USA), were included in the study. The isolates were recovered from patients with bloodstream infections. Susceptibility of the isolates to imipenem, meropenem and ertapenem was detected with microdilution method according to the standards of Clinical and Laboratory Standards Institute (CLSI) minimal inhibitory concentration (MIC) breakpoints. Doripenem susceptibility was detected by the E-test (bioMerieux, Hazelwood, USA). All isolates which were found to be non-susceptible to any of the carbapenem antibiotics tested, were characterized by the phenotypic confirmatory tests and the presence of the resistance genes; blaAmpC, blaCTX-M, blaKPC, blaNDM, blaOXA, blaIMP ve blaVIM were screened by polymerase chain reaction (PCR). Among the 210 ESBL-producing Enterobacteriaceae blood isolates, 23 (11%) were identified as non-susceptible to any of the carbapenems tested. Resistance rates for imipenem, meropenem and ertapenem were 5.7% (n= 12), 1.9% (n= 4) and 2.4% (n= 5), respectively. Doripenem was more active than the other carbapenems, with a resistance rate of 1.0%. Seven of 23 isolates were ESBL negative with cefotaxime/clavulanic acid (CTX/CLA) and ceftazidime/clavulanic acid (CAZ/CLA) combined disk diffusion test, however, six of them were ESBL positive with the addition of boronic acid (BA) to CTX/CLA. Among the three isolates positive for Modifiye Hodge test (MHT) and/or ertapenem-BA tests, blaOXA-48 was detected in one and blaAmpC in the other. Phenotypic pAmpC activity was present in three K.pneumoniae isolates of which one was positive for blaAmpC gene. One K.pneumoniae isolate resistant to all carbapenems with MICs > 256 µg/ml and positive for phenotypic meropenem-BA, MHT, imipenem-EDTA, ceftazidime-CAZ/CLA, cefoxitin-BA production, was found to inhabit blaOXA-48 gene. Five isolates were positive for blaOXA-1 and one for blaOXA-10. Two isolates were positive for blaCTX-M, however blaIMP, blaVIM and blaNDM-1 genes were not detected among the isolates. In conclusion, carbapenem non-susceptibility which was low among the Enterobacteriaceae strains isolated in our center, was mostly attributed to the presence of blaOXA type carbapenemases and no accumulation of blaKPC and blaNDM were detected.
本研究旨在通过表型和基因型方法,调查2005年至2009年间在土耳其安卡拉哈杰泰佩大学医院从侵袭性感染中分离出的肠杆菌科细菌中碳青霉烯类耐药性的存在情况。本研究共纳入210株非重复的大肠杆菌(n = 153)、肺炎克雷伯菌(n = 47)和产酸克雷伯菌(n = 10),这些菌株经BD Phoenix自动鉴定和药敏系统(美国斯帕克斯)检测均确定为超广谱β-内酰胺酶(ESBL)阳性。这些菌株均从血流感染患者中分离获得。根据临床和实验室标准协会(CLSI)最低抑菌浓度(MIC)断点标准,采用微量稀释法检测菌株对亚胺培南、美罗培南和厄他培南的敏感性。采用E-test法(美国海兹伍德生物梅里埃公司)检测多利培南敏感性。所有对所检测的任何一种碳青霉烯类抗生素不敏感的菌株,均通过表型确证试验和耐药基因进行鉴定;通过聚合酶链反应(PCR)筛选blaAmpC、blaCTX-M、blaKPC、blaNDM、blaOXA、blaIMP和blaVIM基因。在210株产ESBL的肠杆菌科血流分离株中,23株(11%)被鉴定为对所检测的任何一种碳青霉烯类抗生素不敏感。亚胺培南、美罗培南和厄他培南的耐药率分别为5.7%(n = 12)、1.9%(n = 4)和2.4%(n = 5)。多利培南比其他碳青霉烯类抗生素活性更高,耐药率为1.0%。23株分离株中有7株通过头孢噻肟/克拉维酸(CTX/CLA)和头孢他啶/克拉维酸(CAZ/CLA)联合纸片扩散试验检测为ESBL阴性,然而,其中6株在CTX/CLA中加入硼酸(BA)后检测为ESBL阳性。在3株改良Hodge试验(MHT)和/或厄他培南-BA试验阳性的分离株中,1株检测到blaOXA-48,另1株检测到blaAmpC。3株肺炎克雷伯菌分离株存在表型pAmpC活性,其中1株blaAmpC基因阳性。发现1株肺炎克雷伯菌分离株对所有碳青霉烯类抗生素耐药,MIC>256 μg/ml,表型美罗培南-BA、MHT、亚胺培南-EDTA、头孢他啶-CAZ/CLA、头孢西丁-BA检测均为阳性,该菌株携带blaOXA-48基因。5株分离株blaOXA-1阳性,1株blaOXA-10阳性。2株分离株blaCTX-M阳性,然而在分离株中未检测到blaIMP、blaVIM和blaNDM-1基因。总之,在我们中心分离的肠杆菌科菌株中,碳青霉烯类不敏感性较低,主要归因于blaOXA型碳青霉烯酶的存在,未检测到blaKPC和blaNDM的积累。