多重耐药肺炎克雷伯菌耐药基因的初步定位研究  

Preliminary locating study on the resistant genes of multi-resistant Klebsiella pneumoniae

作　　者：陈求刚[1] 李银珍[2] 周文[2] 罗兰[1] 

机构地区：[1]中山大学附属第一医院,广东广州510080 [2]中山大学医学院,广东广州510089

出　　处：《中国热带医学》2007年第3期337-338,346,共3页China Tropical Medicine

摘　　要：目的对临床分离的多重耐药肺炎克雷伯菌的耐药基因进行初步定位,为制定临床治疗策略提供理论基础。方法用法国Bio-Merieux公司vitek-60全自动细菌鉴定仪鉴定菌种,双纸片法检测产超广谱β-内酰胺酶,纸片法做药敏试验。用Qiagen plasmid midi kit试剂盒提取该菌质粒,并将质粒电转入感受态大肠杆菌中,用含Amp(50μg/ml)的(LB)选择培养基筛选转化子,然后提取转化子质粒,与肺炎克雷伯菌质粒同时用0.5%的琼脂糖进行电泳分析。肺炎克雷伯菌,电转前、电转后大肠杆菌做药敏试验。结果质粒电泳图谱显示,转化子DH5α所含质粒与肺炎克雷伯菌所含质粒相同,电转化取得成功;肺炎克雷伯菌对17种抗生素均耐药,感受态大肠杆菌对17种抗生素均敏感,转化子DH5α对头孢呋辛和氨苄西林耐药,对其余15种抗生素表现不同程度的敏感。结论本株肺炎克雷伯菌的质粒上可以确定有耐头孢呋辛和氨苄西林的基因,并且能在转化子DH5α中成功表达;头孢噻肟、头孢曲松、头孢哌酮、头孢吡肟、氨曲南的耐药基因可能位于质粒上,但未能完全表达;而环丙沙星、阿米卡星、庆大霉素的耐药基因比较复杂,初步判断不在质粒上;头孢西丁、美洛培南、亚胺培南则无法做出判断。Objective To preliminarily locate the resistant genes of multi - resistant KlebsieUa pneumoniae clinically isolated and provide evidence for reasonable use of antimicrobial agents. Methods The strains were identified by rapid identification testing system of the VITEK-60 SYSTEM（Bio- Merieux） after reactivated, producing extended- spectrumβ- laetamase was detected by double disc synergy test, antumicrobial resistsnce were determined by disk agar diffusion test. Abstracted Klebsiella pneumonia, pinsraids with Qiagen plasmid midi kit and electroporated into Bacillus coll. Plasmids were screened in selected agar （LB） contained AMP （50μg/m]）, abstracted plasmids were analyzed by electrophoresis on the 0.5 % MH agar with Klebsiella pneumoniae plasmids. Results Electropherogram showed that the plasmids in DH5α were the same as those in Klebsiella pneumonia and electroporation was successful; Klebsiella pneumoniae was resistant to 17 antibiotics and Bacillus coil was sensitive to all antibiotics, DH5α was resistant to cefuroxime and ampicillin, but sensitive to others. Conclusion The genes resistant to cefuroxime and ampicillin are located in kpn plasmids and suocefully expressed in DH5α, genes resistant to cefotaxime, eefinaxone, cefoperazone, cefepime, aztreonan may located in kpn plasmid but failed to express. The genes resistant to eiprofloxacin, amikacin, gentamycin are preliminarily considered to be in KlebsieUa pneumoniae pLasmids for their complexity, while location of genes resistant to ceftatine , meropenem, imipenem is so far unknown.

关 键 词：肺炎克雷伯菌 耐药性 基因 

分 类 号：R378.99[医药卫生—病原生物学]