KPC-2基因在克雷伯菌属中传播机制研究

发布时间：2019-06-17 17:21

【摘要】：目的研究克雷伯菌属对亚胺培南耐药机制以及KPC-2基因在克雷伯菌属中传播机制。方法收集四川大学华西医院两个阶段(2009~2010年和2012~2013年)分离的对亚胺培南耐药的克雷伯菌属细菌。琼脂稀释法测定亚胺培南最低抑菌浓度(MIC),CARB ChromID平板和改良Hodges试验检测碳青霉烯耐药表型,PCR检测细菌的KPC-2基因表达。质粒接合试验检测质粒传播性。随机引物扩增多态性DNA(RAPD)方法和肠杆菌基因间重复一致序列聚合酶链反应(ERIC-PCR)技术分别用于分析质粒和菌株的同源性。结果第一阶段筛选并确证耐亚胺培南的3株产酸克雷伯菌首先获得碳青酶烯类抗生素耐药性,第二阶段筛选并确证耐亚胺培南的7株肺炎克雷伯菌出现相同的获得性耐药。PCR显示10株细菌均携带KPC-2型基因。质粒接合试验显示产酸克雷伯菌中携带KPC-2基因的质粒可以传递到受体菌,且与KPC-2基因阳性的肺炎克雷伯菌中携带的质粒具有同源性。ERIC-PCR结果显示7株KPC-2基因阳性的肺炎克雷伯菌具有同源性。结论四川大学华西医院分离的对亚胺培南耐药的肺炎克雷伯菌和产酸克雷伯菌主要耐药机制是产生KPC-2型碳青霉烯酶。产酸克雷伯菌中携带KPC-2基因质粒,该质粒具有传递性且与肺炎克雷伯菌携带质粒相同。肺炎克雷伯菌中耐药株的传播形式为同一克隆传播,而在克雷伯菌属中不同菌种间耐药传播途径为同一质粒的水平传播。
[Abstract]:Objective To study the mechanism of the resistance of Klebsiella to imipenem and the mechanism of the KPC-2 gene in the genus Klebsiella. Methods The isolates of Imipenem-resistant Klebsiella bacteria isolated from the two stages (2009-2010 and 2012-2013) of the West China Hospital of Sichuan University were collected. The minimal inhibitory concentration (MIC) of imipenem, the CARB ChromeID plate and the modified Hodges test were used to detect the resistance phenotype of carbapenem, and the expression of the KPC-2 gene of the bacteria was detected by PCR. Plasmid-binding assay was used to detect plasmid propagation. RAPD and ERIC-PCR were used to analyze the homology of plasmid and strain. Results Three strains of Klebsiella pneumoniae resistant to imipenem were first screened and confirmed to be resistant to carbapenem antibiotics. The second stage of screening and confirmation of imipenem-resistant 7 strains of Klebsiella pneumoniae showed the same acquired resistance. PCR showed that 10 strains of bacteria carried the KPC-2 type gene. The plasmid-binding assay shows that the plasmid carrying the KPC-2 gene in the acid-producing klebsiella can be transferred to the recipient bacterium and has homology with the plasmid carried in the klebsiella pneumoniae positive by the KPC-2 gene. The results of ERIC-PCR show that the 7 KPC-2 gene-positive Klebsiella pneumoniae has homology. Conclusion The main drug-resistant mechanism of Klebsiella pneumoniae and klebsiella for imipenem-resistant Klebsiella pneumoniae isolated from West China Hospital of Sichuan University is the production of the KPC-2 carbapenem. The klebsiella-producing klebsiella carries the KPC-2 gene plasmid, and the plasmid has the transmission property and is the same as that of the Klebsiella pneumoniae carrying plasmid. The transmission forms of the drug-resistant strains in Klebsiella pneumoniae are the same and the transmission of drug-resistant strains in the genus Klebsiella is the same as that of the same plasmid.
【作者单位】：四川大学华西医院实验医学科;
【基金】：国家自然科学基金(No.81000712)资助
【分类号】：R440

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