Detection of Genes of Efflux Pumps (adeB, adeJ and adeG) in Tigecycline Resistant Acinetobacter baumannii Isolated from Benha University Hospital

ALShaimaa ALTabbakh, Sara Saied, Amany K Shahat, Marwa ELMelouk
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Abstract

Background : Emerging of tigecycline resistant Acinetobacter baumannii (A. baumannii) is a critical global health problem as tigecycline is considered the last-line antibiotic for treatment of carbapenem resistant A. baumannii infections. Overexpression of efflux pumps is a leading mechanism of antibiotic resistance in A. baumannii. Objectives: Detection of the presence of three efflux pump genes; adeB, adeJ and adeG and determination of their expression level in Tigecycline Resistant A. baumannii collected from Benha University Hospital. Methodology: Thirty A. baumannii strains were collected and tested for antibiotic susceptibility. Presence of adeB, adeJ and adeG genes was detected by conventional PCR and their expression levels were assessed by RT-PCR. Results: Tigecycline susceptibility showed 60% (18/30) resistance and 40% (12/30) sensitivity. In tigecycline resistant strains, adeB gene was identified in 13/18 (72.2%) and adeJ gene in 12/18 (66.7%) with statistically significant association (P= 0.023). In tigecycline sensitive isolates adeB and adeJ genes were identified in 7/12 (58.3%) and 2/12 (16.7%) strains respectively. The adeG gene was not identified in any of the isolated strains. Combined adeB and adeJ genes were identified in 50% of tigecycline resistant isolates and only in 8.3% of (tigecycline sensitive) isolates with statistical significance difference (P= 0.018). Comparing adeB and adeJ gene expression revealed that transcription level was increased significantly and associated with tigecycline resistance. Conclusion: This study revealed wide spread of tigecycline resistant A. baumannii strains in Benha University Hospital and elucidated the significant role of (adeB and adeJ) genes in their emergence.
本哈大学医院耐替加环素鲍曼不动杆菌外排泵(adeB、adeJ和adeG)基因的检测
背景:耐替加环素鲍曼不动杆菌(鲍曼不动杆菌)的出现是一个严重的全球卫生问题,因为替加环素被认为是治疗耐碳青霉烯类鲍曼不动杆菌感染的最后一线抗生素。外排泵的过度表达是鲍曼不动杆菌耐药的主要机制。目的:检测三种外排泵基因的存在;本哈大学医院采集的耐替加环素鲍曼不动杆菌中adeB、adeJ和adeG的表达及水平测定。方法:收集30株鲍曼不动杆菌进行药敏试验。常规PCR检测adeB、adeJ和adeG基因的存在,RT-PCR检测其表达水平。结果:替加环素药敏率为60%(18/30),敏感性为40%(12/30)。在替加环素耐药菌株中,adeB基因在13/18株中检出(72.2%),adeJ基因在12/18株中检出(66.7%),相关性有统计学意义(P= 0.023)。在替加环素敏感株中,分别在7/12(58.3%)和2/12(16.7%)中检出adeB和adeJ基因。在所有分离菌株中均未发现adeG基因。50%的替加环素耐药菌株检测到adeB和adeJ联合基因,8.3%的替加环素敏感菌株检测到adeB和adeJ基因,差异有统计学意义(P= 0.018)。比较adeB和adeJ基因表达,发现转录水平显著升高,且与替加环素耐药相关。结论:本研究揭示了Benha大学医院耐替加环素鲍曼不动杆菌的广泛分布,并阐明了(adeB和adeJ)基因在其出现中的重要作用。
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