Application of Multiplex PCR for the Identification of Oxacillinase Genes and Determination of Antibiotic Resistance Pattern in Environmental Isolates of Acinetobacter baumannii in ICU

Anahita Farajzadeh, M. Mirzaee, S. Nanekarani, R. Yari
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Abstract

Background: Acinetobacter baumannii is a common cause of nosocomial infections. A prominent feature of these bacteria is resistance to carbapenems. This study aimed to identify OXA genes encoding oxacillinase in Acinetobacter baumannii isolates. Methods: This cross-sectional descriptive study was performed on 25 environmental A. baumannii isolates collected from ICU over 8 months. Definitive identification of isolates was performed by biochemical tests and polymerase chain reaction (PCR) of 16s rRNA gene. Antibiotic susceptibility testing was performed on Müller-Hinton agar medium by disk diffusion and E-test. Antibiogram and multiplex PCR data of beta-lactamase genes were collected and analyzed at a significance level of P<0.05 using SPSS 22.0. Results: Except for one isolate, all isolates (96%) were sensitive to polymyxin B and 80% of isolates were sensitive to oxacillin. All isolates were sensitive to meropenem, ampicillin/sulbactam, gentamicin, amikacin, piperacillin, and carbenicillin. The results showed that 25 isolates (100%) had OXA-51 gene, 21 isolates (84%) had OXA-58 gene, one isolate (4%) had OXA-24 gene, and none of the isolates contained OXA-23 gene. Only isolate No.10 had three oxacillinase genes simultaneously and it was resistant to oxacillin, polymyxin B, and cephalothin. Conclusions: The study showed that environmental isolates of ICU do not have pathogenic genes present in the clinical isolates, and how these genes are transferred to the peripheral isolates is an important point that should be studied. Identification of genes encoding carbapenem resistance may help to understand the mechanisms of resistance transfer in A. baumannii. The lack of the OXA-23 gene plays an important role in the susceptibility of isolates to antibiotics and non-emergence of resistant strains.
多重PCR在ICU环境分离鲍曼不动杆菌Oxacillinase基因鉴定及耐药模式测定中的应用
背景:鲍曼不动杆菌是医院感染的常见原因。这些细菌的一个显著特征是对碳青霉烯类具有耐药性。本研究旨在鉴定鲍曼不动杆菌分离株中OXA基因编码的oxacillinase。方法:采用横断面描述性研究方法,对8个月内从ICU采集的25株环境鲍曼不动杆菌进行研究。采用生化试验和16s rRNA基因聚合酶链反应(PCR)对分离株进行鉴定。采用圆盘扩散法和e试验对 ller- hinton琼脂培养基进行药敏试验。收集β -内酰胺酶基因的抗生素谱和多重PCR数据,使用SPSS 22.0软件在P<0.05的显著水平上进行分析。结果:除1株外,所有菌株(96%)对多粘菌素B敏感,80%对恶西林敏感。所有分离株均对美罗培南、氨苄西林/舒巴坦、庆大霉素、阿米卡星、哌拉西林和卡比西林敏感。结果显示,含OXA-51基因的菌株25株(100%),含OXA-58基因的菌株21株(84%),含OXA-24基因的菌株1株(4%),无OXA-23基因的菌株。只有10号菌株同时具有3个oxacillinase基因,且对oxacillin、多粘菌素B和头孢菌素均耐药。结论:本研究表明,ICU环境分离株不存在临床分离株中存在的致病基因,这些基因如何转移到外周分离株中是一个值得研究的重点。碳青霉烯类耐药基因的鉴定有助于了解鲍曼不动杆菌耐药转移的机制。缺乏OXA-23基因在分离株对抗生素的敏感性和不出现耐药菌株中起重要作用。
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