生物被膜大肠埃希菌AmpC酶、超广谱β-内酰胺酶的检测_药学论文.docVIP

生物被膜大肠埃希菌AmpC酶、超广谱β-内酰胺酶的检测_药学论文 生物被膜大肠埃希菌AmpC酶、超广谱β-内酰胺酶的检测_药学论文 【摘要】 目的 探讨大肠埃希菌生物被膜(biofilm，BF)的耐药情况，为临床合理应用抗生素提供理论依据。 方法 应用改进的平板培养法建立大肠埃希菌BF模型，用银染法和扫描电镜观察鉴定。采用改良三维试验法检测超广谱β-内酰胺酶(ESBLs)及AmpC酶。结果 浮游大肠埃希菌(A组)单产AmpC，单产ESBLs酶及同时产ESBLs和AmpC酶的菌株分别为5.0%(2/40)、30.0%(12/40)和12.5%(5/40)；BF大肠埃希菌(B组)单产AmpC酶，单产ESBLs及同时产ESBLs和AmpC酶的菌株分别为10.0%(4/40)、45.0%(18/40)和22.5%(9/40)。对A组和B组的检出率两两分别进行χ2检验，结果均为0.05。产酶有BF大肠埃希菌对10种抗生素(除亚胺培南全部敏感外)的耐药率均较高。结论 BF的形成和产生ESBLs及AmpC酶的协同作用是大肠埃希菌耐药的主要原因之一。 【关键词】 大肠埃希菌； 生物被膜； 超广谱β-内酰胺酶； AmpC酶 ABSTRACT Objective To evaluate the resistance mechanism of Escherichia coli growing in biofilm, and to provide the theoretical guide to select antimicrobial agents in clinic. Methods Model of Escherichia coli bacterial biofilm was built up by the modified flat-board method and identified by the method staining with AgNO3 and confocal scanning laser microscopy. ESBLs and AmpC β-lactamase were detected by improved three-dimension test. Results The detection rates of AmpC, ESBLs and ESBLs plus AmpC β-lactamase Escherichia coli were 5.0% (2/40), 30.0% (12/40) and 12.5% (5/40), whereas those in biofilm Escherichia coli were 10.0% (4/40), 45.0% (18/40) and 22.5% (9/40) respectively. the resistant rate of the biofilm Escherichia coli producing AmpC and ESBLs to 10 kinds of antibiotics was higher (except imipenem). Conclusion One of the main reasons that Escherichia coli was resistant to antibiotics was the synergetic effect of ESBLs, AmpC and biofilm. KEY WORDS Escherichia coli; Biofilm; ESBLs; AmpC enzyme 大肠埃希菌是各类感染常见的致病菌，易于固着在生物材料的表面生长，并形成生物被膜(biofilm，BF)。常规剂量的抗生素治疗无效，常常导致感染迁延不愈和反复急性发作。关于BF菌的耐药机制引起国内外学者的广泛关注，研究表明，大肠埃希菌对β-内酰胺类抗生素耐药主要是由质粒介导的超广谱β-内酰胺酶(extended-spectrum β-lactamases，ESBLs)和持续高水平表达的AmpC酶引起的。细菌形成BF后，其产β-内酰胺酶的活性明显增加［1，2］。本实验通过建立大肠埃希菌BF模型，检测BF内AmpC酶及ESBLs的产生情况，并进一步探讨其耐药机制，供临床选择抗生素参考。1 材料和方法 1.1 材料 大肠埃希菌40株(本院细菌室分离并鉴定)，质控菌株为大肠埃希菌ATCC25922、肺炎克雷伯菌ATCC700603、阴沟肠杆菌029及029M。所有菌株均经VITEX自动化微生物分析仪鉴定。培养基胰酶大豆肉汤(TSB)为法国BioMerieux公司产品。主要仪器与药品有医用硅胶膜(1.5mm，北京医用硅胶膜研究所)、2