【摘要】 目的 分析碳青霉烯类耐药肺炎克雷伯菌临床株K27的基因组和携带的质粒的特征。方法 通过VITEK 2 Compact系统（法国梅里埃）测定抗菌药物敏感性。使用S1核酸酶脉冲场凝胶电泳（S1-PFGE）估计质粒大小。通过接合实验评估质粒转移能力。使用Illumina和Oxford Nanopore平台进行全基因组测序（WGS）。利用生物信息学方法分析基因组特征。结果 菌株K27是一种泛耐药ST11型肺炎克雷伯菌，能产生3种碳青霉烯酶（NDM-5、KPC-2和VIM-1）。S1-PFGE和全基因组测序均显示，K27菌株携带4个质粒。其中，碳青霉烯酶基因bla_KPC-2位于IncFII（pHN7A8）/IncR杂合质粒（pK27-KPC）上，而碳青霉烯酶基因bla_NDM-5和bla_VIM-1基因由同一IncA质粒（pK27-NDM/VIM）携带。液相接合实验表明pK27-NDM/VIM为接合质粒。结论 这是首次关于临床分离的ST11肺炎克雷伯菌K27携带的碳青霉烯酶基因bla_NDM-5和bla_VIM-1位于同一个质粒上的报道。共携带多种碳青霉烯酶基因和多种类抗菌药物耐药基因使该菌株呈现泛耐药，从而对临床治疗构成严重威胁。更多还原

【Abstract】 Objective To analyze the genome and the characteristics of the plasmids carried by the carbapenem-resistant Klebsiella pneumoniae clinical strain K27.Methods Antimicrobial susceptibility testing was determined by the VITEK 2 Compact system（bioMérieux, France）.The sizes of the plasmid was estimated by S1 nuclease pulsed-field gel electrophoresis（S1-PFGE）.The ability of plasmid transfer was evaluated by conjugation experiments.Whole genome sequencing（WGS） was performed using the Illumina and Oxford Nanopore platforms.Genomic characteristics were analyzed using bioinformatics methods.Results Strain K27 is a pan-resistant ST11 Klebsiella pneumoniae that can produce three types of carbapenemases（NDM-5,KPC-2 and VIM-1）.Both S1-PFGE and whole-genome sequencing revealed that the stain K27 carried four plasmids.Among them, the carbapenemase gene bla_KPC-2 was located on the IncFII（pHN7A8）/IncR hybrid plasmid（pK27-KPC）,while the carbapenemase genes bla_NDM-5 and bla_VIM-1 were carried by the same IncA plasmid（pK27-NDM/VIM）.The conjugation experiments indicated that pK27-NDM/VIM was a conjugative plasmid.Conclusion This is the first report of a plasmid carrying both the carbapenemase genes bla_NDM-5 and bla_VIM-1 in a clinical ST11 Klebsiella pneumoniae strain K27.The presence of multiple carbapenemase genes and multiple types of antimicrobial resistance genes in this strain makes it highly pan-resistant, thereby presenting a serious threat to treatment in clinical practice.更多还原