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目的了解云南省昆明某医院临床铜绿假单胞菌分离株耐药特征及耐药基因分子流行病学特征,为临床治疗与控制院内感染提供依据。方法 2013年6 9月共收集全院不同病区感染患者28株铜绿假单胞菌。微生物自动分析仪鉴定菌株及耐药分析。PCR扩增及序列分析β-内酰胺酶bla基因、氟喹诺酮类及氨基糖苷类耐药基因及IS插入元件及整合子。脉冲场凝胶电泳(pulsed field gel electrophoresis,PFGE)分析菌株间遗传变异关系。结果全部菌株呈现多重耐药且存在有广泛耐药性(extensively drug-resistant,XDR)及泛耐药性(pendrug-resistant,PDR)菌。100%(28/28)菌株对一代/二代及大部分三代/四代头孢霉素、磺胺类及叶酸代谢途径抑制剂耐药。85.7%(24/28)菌株对碳青霉烯类抗生素美罗培南(MEM)、亚胺培南(IMP)耐药,50.0%(14/28)菌株对厄他培南(ETP)耐药。92.9%(26/28)菌株同时携带β-内酰胺酶基因、氟喹诺酮类及氨基糖苷类耐药基因,且菌株间基因谱存在较大不同。blaFOX/qnr B/aac A4为主要耐药基因谱(21.4%,6/28)。57.1%(16/28)菌株携带int1整合酶基因,21.4%(6/28)菌株携带ISCR1插入元件,35.7%(10/28)菌株携带ISEcp1插入元件。PFGE显示14株菌株分为8个克隆群。结论本研究铜绿假单胞菌分离株存在较高耐药率,多种耐药基因存在不同克隆菌株中,提示耐药基因在菌株间及不同菌种间存在快速传播风险。
Objective To understand the drug resistance characteristics and the molecular epidemiological characteristics of clinical isolates of Pseudomonas aeruginosa isolated from a hospital in Kunming, Yunnan Province, and provide the basis for clinical treatment and control of nosocomial infections. Methods A total of 28 strains of Pseudomonas aeruginosa were collected from patients with different wards in the hospital from June to September 2013. Microbial Identification of strains and automatic analysis of drug resistance. PCR amplification and sequence analysis of β-lactamase bla genes, fluoroquinolones and aminoglycoside resistance genes and IS insertion elements and integrons. Pulsed field gel electrophoresis (PFGE) analysis of genetic variation among isolates. Results All strains showed multidrug resistance and there was extensively drug-resistant (XDR) and pendrug-resistant (PDR) bacteria. 100% (28/28) strains were resistant to first / second and most third / fourth generation cephalosporins, sulfonamides and folic acid metabolic pathway inhibitors. 85.7% (24/28) strains were resistant to carbapenem meropenem (MEM) and imipenem (IMP), and 50.0% (14/28) strains were resistant to ertapenem (ETP). 92.9% (26/28) of the strains carried both β-lactamase gene, fluoroquinolone and aminoglycoside resistance genes, and there was a big difference in the gene spectrum between strains. blaFOX / qnr B / aac A4 is the major drug resistance gene spectrum (21.4%, 6/28). 57.1% (16/28) of the strains carried the int1 integrase gene, 21.4% (6/28) of the strains carried the ISCR1 insert, and 35.7% (10/28) carried the ISEcp1 insert. PFGE showed 14 strains were divided into 8 clonal groups. Conclusion There is a high resistance rate of Pseudomonas aeruginosa isolates in this study, and many resistant genes exist in different clonal strains, suggesting that there is a risk of rapid spread of drug-resistant genes among strains and different strains.