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中国镇江分离的多重耐药鲍曼不动杆菌中与耐药相关的β-内酰胺酶、膜孔蛋白和外排泵基因的特征及分布

Characterization and distribution of drug resistance associated β-lactamase, membrane porin and efflux pump genes in MDR A. baumannii isolated from Zhenjiang, China.

作者信息

Yang Huijian, Huang Lan, Barnie Prince Amoah, Su Zhaoliang, Mi Zuhuang, Chen Jianguo, Aparna Vasudevan, Kumar Dinesh, Xu Huaxi

机构信息

Department of Immunology, School of Medicine, Jiangsu University Zhenjiang 212013, PR China.

Wuxi Clone Gen-Tech Institute Wuxi 214026, PR China.

出版信息

Int J Clin Exp Med. 2015 Sep 15;8(9):15393-402. eCollection 2015.

Abstract

BACKGROUND

Acinetobacter baumannii (A. baumannii), especially the multidrug resistant A. baumannii (MDR-AB) is becoming a common opportunistic pathogen in hospital, and constitutes significant public health threats. This study aimed at investigating the relationship between drug resistance with expression of class A-D β-lactamase genes, mutation in membrane porin and over-expression of efflux pump genes among A. baumannii isolated from Zhengjiang, China.

METHODS

Antibiotic susceptibility assays were performed using Kirby-Bauer disc diffusion method. PCR was used to detect β-lactamase genes and carO, oprD, adeR, adeS. Real-time PCR was used to assess the mRNA expression level of efflux pump gene adeB. The software of DNAMAN was applied to assemble oprD and carO sequences, and the sequences were compared with those retrieved from GenBank (http://www.ncbi.nlm.nih.gov/).

RESULTS

27 isolates (61.4%) in this study were MDR-AB, in which five β-lactamases including TEM, CTX-M-2, ADC, OXA-23 and OXA-51 were found, and the positive rate was 96.3% (26), 14.8% (4), 92.6% (25), 88.9% (24) and 92.6% (25), respectively. In addition, the expression level of adeB mRNA was significantly increased in MDR-AB, it might due to adeR mutation. Some mutations were also found in carO and oprD.

CONCLUSION

MDR-AB showed high relationship with β-lactamase, mutation in membrane porin and overexpression of adeB, which may directly relates to the mutation in regulating gene adeR.

摘要

背景

鲍曼不动杆菌,尤其是多重耐药鲍曼不动杆菌(MDR-AB)正成为医院中常见的机会致病菌,对公共卫生构成重大威胁。本研究旨在调查从中国镇江分离的鲍曼不动杆菌中耐药性与A-D类β-内酰胺酶基因表达、膜孔蛋白突变及外排泵基因过表达之间的关系。

方法

采用 Kirby-Bauer 纸片扩散法进行抗生素敏感性试验。用 PCR 检测β-内酰胺酶基因及 carO、oprD、adeR、adeS。用实时 PCR 评估外排泵基因 adeB 的 mRNA 表达水平。应用 DNAMAN 软件组装 oprD 和 carO 序列,并与从 GenBank(http://www.ncbi.nlm.nih.gov/)检索到的序列进行比较。

结果

本研究中 27 株分离菌(61.4%)为 MDR-AB,其中发现了包括 TEM、CTX-M-2、ADC、OXA-23 和 OXA-51 在内的五种β-内酰胺酶,阳性率分别为 96.3%(26)、14.8%(4)、92.6%(25)、88.9%(24)和 92.6%(25)。此外,MDR-AB 中 adeB mRNA 的表达水平显著升高,这可能是由于 adeR 突变所致。在 carO 和 oprD 中也发现了一些突变。

结论

MDR-AB 与β-内酰胺酶、膜孔蛋白突变及 adeB 过表达高度相关,这可能直接与调节基因 adeR 的突变有关。

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