Keskin Hakan, Tekeli Alper, Dolapci İştar, Öcal Duygu
Sinop Ataturk State Hospital, Microbiology Laboratory, Sinop, Turkey.
Mikrobiyol Bul. 2014 Jul;48(3):365-76.
Acinetobacter baumannii is an important cause of nosocomial infections that particularly increase the mortality and the morbidity at the intensive care units of the hospitals. The aims of this study were to evaluate the resistance genes, antibiotic susceptibility and the clonal relations among Acinetobacter strains isolated from clinical samples and to determine the resistance mechanisms related to these bacteria in our hospital. A total of 201 A.baumannii strains isolated from different clinical samples (35.3% from tracheal aspirate, 27.3% from blood, 18.4% from abscess material, 19% from other samples) of 160 inpatients evaluated at the Ibni Sina Hospital Central Bacteriology Laboratory, Ankara University School of Medicine, Turkey from April 2010 to December 2011, were included in the study. Identification of the isolates and their susceptibility testing against amikacin, ciprofloxacin, tetracycline, sulbactam/ampicillin, trimethoprim/sulfametoxazole (SXT), ceftazidime, gentamicin, imipenem, levofloxacin, meropenem, piperacillin/tazobactam, cefoperazone/sulbactam, cefepime and colistin were performed by the automated systems, namely Vitek 2 (bioMérieux, France) and BD Phoenix (Becton Dickinson, USA). The molecular mechanisms of beta-lactamase resistance and the presence of integrons were analyzed by polymerase chain reaction (PCR). Moreover, since blaPER-1 gene is of high frequency in Turkey, it was also investigated in the isolates. Pulsed-field gel electrophoresis (PFGE) was performed to examine the clonal relations between isolates. Our results indicated that multidrug resistance rate of A.baumannii was 94.5% (190/201), while 94% (189/201) of the isolates were susceptible to colistin thus making it the most potent antimicrobial agent, followed by amikacin and SXT with a susceptibility rate of 32%. Twelve colistin-resistant isolates were further investigated with the E-test method (AB Biodisk, Sweden) and found to be colistin-resistant. While the results were negative for the genes responsible from metallo-beta-lactamase production, positive results were obtained for blaOXA genes at various rates (OXA-51 100%; OXA-23 91.5%; OXA-58 7%; OXA-24 2%). PFGE results revealed four different main clones (29 isolates in genotype A, 23 in genotype B, 18 in genotype C and 7 in genotype D) in the study population. No common epidemic isolate was detected. Class 1 integrons which take part in the transfer of resistance genes were detected in 112 (55.7%) isolates. There was no statistically significant difference between the genotype distributions of class 1 integron positive strains (p> 0.05). The relationship between the presence of integron in multidrug resistant isolates and resistance to tetracyclin, SXT, imipenem, meropenem, cefoperazone/sulbactam and cefepime were found to be statistically significant (p< 0.05). Of the isolates 42 (21%) were positive for blaPER- 1 gene and all were resistant to ceftazidime. This study indicated that blaOXA genes found together with blaOXA-51 genes play an important role in carbapenem resistance of A.baumannii strains. Moreover, multidrug resistance is still an important problem in infections caused by A.baumannii and integrons play a role in the transfer of the resistance genes. In conclusion, multidrug resistant A.baumannii strains were common in our hospital and our epidemiologic data would be helpful for further investigations and in therapeutical approaches.
鲍曼不动杆菌是医院感染的重要病因,尤其会增加医院重症监护病房的死亡率和发病率。本研究的目的是评估从临床样本中分离出的不动杆菌菌株的耐药基因、抗生素敏感性及克隆关系,并确定我院这些细菌的耐药机制。2010年4月至2011年12月期间,在土耳其安卡拉大学医学院伊本·西那医院中央细菌学实验室对160名住院患者的不同临床样本(35.3%来自气管吸出物,27.3%来自血液,18.4%来自脓肿材料,19%来自其他样本)进行检测,共分离出201株鲍曼不动杆菌,纳入本研究。采用Vitek 2(法国生物梅里埃公司)和BD Phoenix(美国贝克曼库尔特公司)等自动化系统对分离菌株进行鉴定,并检测其对阿米卡星、环丙沙星、四环素、舒巴坦/氨苄西林、甲氧苄啶/磺胺甲恶唑(SXT)、头孢他啶、庆大霉素、亚胺培南、左氧氟沙星、美罗培南、哌拉西林/他唑巴坦、头孢哌酮/舒巴坦、头孢吡肟和黏菌素的敏感性。通过聚合酶链反应(PCR)分析β-内酰胺酶耐药的分子机制及整合子的存在情况。此外,由于blaPER-1基因在土耳其的频率较高,也对分离菌株进行了检测。采用脉冲场凝胶电泳(PFGE)检测分离菌株之间的克隆关系。我们的结果表明,鲍曼不动杆菌的多重耐药率为94.5%(190/201),而94%(189/201)的分离菌株对黏菌素敏感,因此黏菌素是最有效的抗菌剂;其次是阿米卡星和SXT,敏感性率为32%。采用E-test法(瑞典AB生物盘公司)对12株黏菌素耐药分离株进行进一步检测,结果显示它们对黏菌素耐药。金属β-内酰胺酶产生相关基因的检测结果为阴性,而blaOXA基因的检测结果呈不同比例阳性(OXA-51 100%;OXA-23 91.5%;OXA-58 7%;OXA-24 2%)。PFGE结果显示,研究人群中有四个不同的主要克隆(基因型A中有29株,基因型B中有23株,基因型C中有18株,基因型D中有7株)。未检测到常见的流行菌株。在112株(55.7%)分离菌株中检测到参与耐药基因转移的1类整合子。1类整合子阳性菌株的基因型分布之间无统计学显著差异(p>0.05)。多重耐药分离株中整合子的存在与对四环素、SXT、亚胺培南、美罗培南、头孢哌酮/舒巴坦和头孢吡肟的耐药之间的关系具有统计学意义(p<0.05)。42株(21%)分离菌株的blaPER-1基因呈阳性,且均对头孢他啶耐药。本研究表明,与blaOXA-51基因同时存在的blaOXA基因在鲍曼不动杆菌菌株的碳青霉烯耐药中起重要作用。此外,多重耐药仍是鲍曼不动杆菌引起的感染中的一个重要问题,整合子在耐药基因的转移中起作用。总之,多重耐药鲍曼不动杆菌菌株在我院很常见,我们的流行病学数据将有助于进一步研究和治疗方法的制定。