摘要
高毒力碳青霉烯耐药肺炎克雷伯菌(Hv-CRKP)已成为全球公共卫生领域的重大挑战.该研究报告了2017年至2022年浙江一家三甲医院Hv-CRKP分离株的定植和传播情况.对来自72名患者的90个不同的CRKP菌株进行测序分析,发现院内感染Hv-CRKP的流行主要依赖于ST11-K64克隆传播.对11个代表性分离株进行了全基因组测序,得到了31个完整的质粒基因序列,其中包括12个携带KPC-2耐药基因的质粒和10个携带rmpA毒力基因的质粒.除了二元质粒外,还发现了两种可介导rmpA和KPC-2共同传播的融合质粒.研究人员通过捕获其祖先质粒,提出了该融合质粒的形成机制.此外,发现了五种rmpA启动子变异体(P9T到P13T),其活力与地理分布存在差异.通过CRISPR/Cas9基因编辑技术,证实活跃的“P11T-rmpA”调控子是“高风险”ST11-K64/CRKP克隆群的生物标志物.该研究拓展了对ST11-K64/Hv-CRKP流行性克隆的播散和临床演进的认知.
The spread of hypervirulent carbapenem-resistant Klebsiella pneumoniae(Hv-CRKP)is a global health concern.Here,we report the intrahospital colonization and spread of Hv-CRKP isolates in a tertiary hospital from 2017 to 2022.Analyses of 90 nonredundant CRKP isolates from 72 patients indicated that Hv-CRKP transferability relies on the dominant ST11-K64 clone.Whole-genome sequencing of 11 representative isolates gave 31 complete plasmid sequences,including 12 KPC-2 resistance carriers and 10 RmpA virulence vehicles.Apart from the binary vehicles,we detected two types of fusion plasmids,favoring the cotransfer of RmpA virulence and KPC-2 resistance.The detection of ancestry/relic plasmids enabled us to establish genetic mechanisms by which rare fusion plasmids form.Unexpectedly,we found a total of five rmpA promoter variants(P9T–P13T)exhibiting distinct activities and varying markedly in their geographic distributions.CRISPR/Cas9 manipulation confirmed that an active PT11-rmpA regulator is a biomarker for the"high-risk"ST11-K64/CRKP clone.These findings suggest clonal spread and clinical evolution of the prevalent ST11-K64/Hv-CRKP clones.Apart from improved public awareness of Hv-CRKP convergence,our findings might benefit the development of surveillance(and/or intervention)strategies for the dominant ST11-K64 lineage of the Hv-CRKP population in healthcare sectors.
作者
刘力彰
娄苧洁
梁琦强
肖伟
滕高钦
马剑钢
张会敏
黄曼
冯友军
Lizhang Liu;Ningjie Lou;Qiqiang Liang;Wei Xiao;Gaoqin Teng;Jiangang Ma;Huimin Zhang;Man Huang;Youjun Feng(Key Laboratory of Multiple Organ Failure,Ministry of Education,Department of Microbiology and General Intensive Care Unit of the Second Affiliated Hospital,Zhejiang University School of Medicine,Hangzhou 310058,China;Zhejiang Academy of Agricultural Sciences,Hangzhou 310021,China;Cancer Center at Illinois,University of Illinois at Urbana-Champaign,Urbana,IL 61801,USA;Department of Clinical Laboratory,Shenzhen Third People’s Hospital,National Clinical Research Center for Infectious Diseases,The Second Affiliated Hospital of Southern University of Science and Technology,Shenzhen 518112,China;Zhejiang Provincial Key Laboratory for Microbial Biochemistry and Metabolic Engineering,Hangzhou 310058,China)
基金
supported by the National Natural Science Foundation of China(32141001 and 31830001)
the National Science Fund for Distinguished Young Scholars(32125003)
the National Key R&D Program of China(2022YFC2303900 and 2022YFC3704700)。
