Department of Infectious Disease, The First Affiliated Hospital of Xinxiang Medical University, Xinxiang 453100, P.R. China.
Mol Med Rep. 2013 Feb;7(2):664-8. doi: 10.3892/mmr.2012.1235. Epub 2012 Dec 14.
The aim of the present study was to investigate the resistance of plasmid-mediated AmpC β-lactamase in Pseudomonas aeruginosa, to detect and identify the AmpC genotype and to provide evidence for antibiotic applications in the clinic. Resistance phenotype in 108 strains of clinically isolated P. aeruginosa was determined by Kirby-Bauer disk test and cefoxitin three dimensional test in AmpC-positive strains. Plasmids were extracted from AmpC-positive strains using the SDS-alkali splitting technique. The depurated plasmid was used to amplify AmpC β-lactamase genes by PCR. Positive PCR products were sequenced by the Shanghai Sangon Biological Engineering Technology Company. Gene homology of PCR products with other index sample gene sequences was compared. In the present study, 28 AmpC enzyme-positive P. aeruginosa strains among 108 were identified. Multidrug‑resistance to antibiotics was observed in positive AmpC P. aeruginosa strains and a new P. aeruginosa strain of plasmid-mediated CMY-7 type AmpC enzyme was identified. In addition, AmpC type β-lactamases were revealed to be important in the resistance mechanism to antibiotics in P. aeruginosa. This is the first report of CMY-7 plasmid‑mediated AmpC enzyme expression in P. aeruginosa.
本研究旨在探讨铜绿假单胞菌中质粒介导的 AmpC β-内酰胺酶的耐药性,检测和鉴定 AmpC 基因型,并为临床抗生素应用提供依据。采用 Kirby-Bauer 纸片扩散法和头孢西丁三维试验对 108 株临床分离的铜绿假单胞菌进行耐药表型检测,对 AmpC 阳性菌株采用 SDS-碱裂解法提取质粒。采用 PCR 扩增 AmpC β-内酰胺酶基因,对阳性 PCR 产物进行上海生工生物工程技术服务有限公司测序,比较 PCR 产物与其他指数样本基因序列的同源性。本研究从 108 株菌中鉴定出 28 株 AmpC 酶阳性铜绿假单胞菌。对阳性 AmpC 铜绿假单胞菌进行药敏试验,发现其对多种抗生素耐药,鉴定出一株新的质粒介导的 CMY-7 型 AmpC 酶铜绿假单胞菌。此外,AmpC 型β-内酰胺酶在铜绿假单胞菌对抗生素的耐药机制中起重要作用。这是首次报道铜绿假单胞菌中 CMY-7 质粒介导的 AmpC 酶表达。