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Distribution of Aminoglycoside Resistance Mediated by 16S rRNA Methylation among IraqI Isolates of Escherichia coli and Pseudomonas aeruginosa

机译:大肠杆菌和铜绿假单胞菌伊拉克分离株中由16S rRNA甲基化介导的氨基糖苷抗性分布

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One hundred clinical isolates of Escherichia coli and Pseudomonas aeruginosa (58 , 42 isolates respectively ) were obtained from patients suffering from different infections at Baghdad Iraq teaching hospital . These isolates were diagnosed using api 20E .Results of primary screening test for aminoglycoside resistance using determination the minimum inhibitory concentration revealed that all the isolates conferring multidrug resistance and the highest resistance was against kanamycin, while the lowest was against amikacin .Phenotypic detection of Extended spectrum ?–lactamase ( ES?Ls) was preformed and the results showed that 84% of the isolates gave positive results. Highly resistant isolates (20 for each ) were selected for the genetic study using polymerase chain reaction technique (PCR) to determine aminoglycoside resistance mediated by methylation 16S rRNA beside detection blaCTX –M gene responsible for ESBLs production .Seven 16S rRNA methylase genes were amplified ,the ArmA (846 bp), RmtA(635bp), RmtB(584bp), RmtC(711bp), RmtD (500 bp), RmtF(453bp) and npmA (641bp) beside amplifying blaCTX –M gene ( 550bp) .Out of 20 E.coli isolates ,16(80%)gave positive results for ArmA gene, while non of P.aeruginosa harboured this gene. Only one isolates out of 20(5%) harboured RmtB methylation gene in E.coli isolates, while 3 isolates out of 20(15%) contains RmtC gene and 1 isolates(5%) harboured RmtD gene in E.coli isolates while in P.aeruginosa showed 3 isolates out of 20 (15%) positive results in this gene. The sixth methylation gene was npmA was detected in only one isolate (5%) out of 20. For blaCTX –M gene , it was detected in all E.coli isolates (100%) while it was detected in 17(85%) of P.aeruginosa. This is the first report in Iraq for the emergence of 16S rRNA methylases among E.coli and P.aeruginosa in correlation with ES?Ls production . Key words: Aminoglycoside resistance, 16S rRNA methylation genes, and ES?Ls blaCTX –M genes.
机译:在巴格达伊拉克教学医院从患有不同感染的患者中获得了一百株临床分离的大肠杆菌和铜绿假单胞菌(分别为58株和42株)。这些分离物使用api 20E诊断。通过确定最小抑菌浓度的氨基糖苷抗性的初步筛选测试结果显示,所有分离物均具有多药耐药性,对卡那霉素的耐药性最高,而对阿米卡星的耐药性最低。扩展谱的表型检测预先制备了β-内酰胺酶(ESβLs),结果表明84%的分离物给出了阳性结果。使用聚合酶链反应技术(PCR)选择高抗性菌株(每个20株)进行遗传研究,以确定由甲基化16S rRNA介导的氨基糖苷抗性,以及负责ESBLs产生的检测blaCTX -M基因。扩增了七个16S rRNA甲基化酶基因, ArmA(846 bp),RmtA(635bp),RmtB(584bp),RmtC(711bp),RmtD(500 bp),RmtF(453bp)和npmA(641bp)以及blaCTX –M基因(550bp)的扩增。大肠杆菌分离物,16(80%)对ArmA基因有阳性结果,而非P. aruginosa则含有该基因。在大肠杆菌中,只有20(5%)的分离株带有RmtB甲基化基因,而在大肠杆菌中,只有20(15%)的3个分离株含有RmtC基因,而在大肠杆菌中有1(5%)的RmtD基因带有RmtD基因。铜绿假单胞菌在该基因的20个阳性结果中显示3个分离株(15%)。第六个甲基化基因是npmA,仅在20个菌株中检出一个(5%)。对于blaCTX –M基因,在所有大肠杆菌分离株(100%)中都检出,而在17个(85%)大肠杆菌中检出。铜绿假单胞菌这是伊拉克关于大肠杆菌和铜绿假单胞菌中与ES?Ls产生有关的16S rRNA甲基化酶的首次报道。关键词:氨基糖苷抗性,16S rRNA甲基化基因和ES?Ls blaCTX –M基因。

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