肺炎克雷伯菌OqxAB外排泵与生物膜形成相关性研究

发布时间：2018-06-23 21:37

本文选题：肺炎克雷伯菌 + 生物膜　；参考：《中国抗生素杂志》2017年08期

【摘要】：目的分析耐碳青霉烯类肺炎克雷伯菌(CRKPs)的生物膜形成能力与OqxAB外排泵的相关性。方法收集2008—2013年间温州医科大学附属第一医院临床分离的碳青霉烯类耐药肺炎克雷伯菌37株。采用微量肉汤稀释法检测其对亚胺培南、厄他培南以及美罗培南的最低抑菌浓度(minimal inhibitory concentration,MIC),结晶紫染色法测定其生物膜形成能力,PCR方法筛查外排泵基因oqxA和oqxB的携带情况并分析亚抑菌浓度的外排泵抑制剂羰基氰氯苯腙(CCCP)对生物膜形成的影响;实时荧光定量PCR(qRT-PCR)检测CCCP存在与不存在条件下生物膜状态与浮游状态oqxB外排泵基因的表达量。用统计学软件SPSS 17.0分析实验数据。结果 37株肺炎克雷伯菌均对至少1种碳青霉烯类药物的敏感性降低,37株菌都具有不同程度的生物膜形成能力(A590范围为0.044~1.113);在所有菌株中,只有7株菌存在oqxA或oqxB外排泵基因的缺失,与外排泵基因完整的试验菌株相比,这7株菌形成生物膜的能力较低;外排泵抑制试验发现亚抑菌浓度的CCCP对实验菌株的生物膜形成能力都有不同程度的促进作用;qRT-PCR结果证明,生物膜状态下菌株的外排泵基因的表达量要高于浮游状态下的表达量。结论 OqxAB外排泵过表达可能会促进肺炎克雷伯菌生物膜的形成。
[Abstract]:Objective to analyze the relationship between biofilm forming ability of Klebsiella pneumoniae (CRKPs) and OqxAB efflux pump. Methods 37 strains of carbapenem resistant Klebsiella pneumoniae isolated from the first affiliated Hospital of Wenzhou Medical University from 2008 to 2013 were collected. Determination of imipenem by broth dilution method, The minimum inhibitory concentrations of ertapenem and meropenem (minimal inhibitory concentration), the biofilm formation ability of ertapenem and meropenem were determined by crystal violet staining. PCR was used to screen for the carrying of efflux pump gene oqxA and oqxB, and to analyze the carbonyl of efflux pump inhibitor of subinhibitory concentration. The effect of CCCP on the formation of biofilm; Real-time fluorescence quantitative PCR (qRT-PCR) was used to detect the expression of oqxB efflux pump gene in the presence and absence of CCCP. Statistical software SPSS 17.0 was used to analyze the experimental data. Results the sensitivity of 37 strains of Klebsiella pneumoniae to at least one carbapenes was decreased. 37 strains of Klebsiella pneumoniae showed different biofilm forming ability (A590 range: 0.044% 1.113). Only 7 strains had the absence of oqxA or oqxB efflux pump genes. Compared with the experimental strains with intact efflux pump genes, the biofilm formation ability of these 7 strains was lower. The results of efflux pump inhibition test showed that CCCP with subinhibitory concentration could promote the biofilm formation ability in different degree. The results of qRT-PCR showed that the expression of efflux pump gene in biofilm was higher than that in planktonic state. Conclusion the overexpression of OqxAB efflux pump may promote the biofilm formation of Klebsiella pneumoniae.
【作者单位】：温州医科大学附属第一医院医学检验中心;浙江医院检验科;温州医科大学检验医学院生命科学学院;
【基金】：浙江省卫生高层次创新人才培养工程项目(No.浙卫发[2012]241号)
【分类号】：R446.5

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