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High Expression of Metallo-β-Lactamase Contributed to the Resistance to Carbapenem in Clinical Isolates of Pseudomonas aeruginosa from Baotou, China

机译:金属-β-内酰胺酶的高表达有助于临床分离株铜绿假单胞菌的抗胰岛蛋白酶患者

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Background: Bacterial resistance to antibiotics has become a major public health concern. This study aimed to determine the resistance mechanisms to carbapenem in clinical isolates of Pseudomonas aeruginosa . Methods: A total of 62 clinical isolates of carbapenem-resistant P. aeruginosa (CRPA) were collected from 2015 to 2017. Imipenem (IPM)–EDTA disk synergy test was used to screen strains that produced metallo-β-lactamase. In addition, the genes for outer membrane protein OprD2, metallo-β-lactamase and mexR gene were amplified and sequenced. Expression of mexA was detected by real-time PCR. Results: Disk synergy test showed that 51.6% (32/62) of the strains were positive for metallo-β-lactamase. PCR showed that 84.4% of the strains were SIM-positive (27/32), 15.6% of the strains were IMP-positive (5/32), and 12.5% of the strains were VIM-positive (4/32). SPM-positive and GIM-positive strains were not detected. In addition, 5 of the 62 strains had small deletions and/or point mutations in OprD2. Three strains had a high expression of mexA, while eight strains were positive for the regulatory gene mexR with no mutations detected by DNA sequencing. Conclusion: Expression of metallo-β-lactamase is the main resistance mechanism of P. aeruginosa to carbapenem. Mutations in OprD2 and/or the overexpression of efflux pump MexAB-OprM may contribute to P. aeruginosa resistance to carbapenem.
机译:背景:对抗生素的细菌抗性已成为主要的公共卫生问题。本研究旨在确定铜绿假单胞菌临床分离株患者对Carbapenem的抵抗机制。方法:从2015〜2017年收集了耐抵抗铜绿假单胞菌P.铜绿假单胞菌(CRPA)的62分离株。Imipenem(IPM)-edTA盘协同作用试验用于筛选产生金属-β-内酰胺酶的菌株。此外,扩增和测序外膜蛋白OPRD2,Metallo-β-内酰胺酶和MEXR基因的基因。通过实时PCR检测MEXA的表达。结果:磁盘协同试验表明,51.6%(32/62)的菌株对于金属-β-内酰胺酶是阳性的。 PCR显示,84.4%的菌株是SIM阳性(27/32),15.6%的菌株是灭毒(5/32),12.5%的菌株是Vim阳性(4/32)。没有检测到SPM阳性和GIM阳性菌株。此外,62个菌株中的5个具有小缺失和/或OPRD2中的突变。三种菌株具有高表达Mexa,而八种菌株对于监管基因MEXR是阳性的,没有通过DNA测序检测的突变。结论:金属-β-内酰胺酶的表达是铜绿假单胞菌至肉豆蔻蛋白的主要耐药机制。 OPRD2和/或Efflux泵Mexab-OPRM的过表达的突变可能有助于铜绿假单胞菌对Carbapenem的抗性。

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