摘要
目标
blaKPC-2在医院环境中的出现已成为世界范围内的一场重大公共卫生危机。我们的目的是对从住院患者中分离的三株产KPC的革兰氏阴性杆菌（KPC-GNB）进行全基因组测序（WGS），以鉴定获得性抗微生物耐药性基因（ARGs）。
方法
使用Illumina MiSeq-I进行WGS，并使用SPAdes实现从头组装。使用Resfinder、AMRFinder、ISFinder、plasmidSPAdes、PlasmidFinder、MOB套件、PLSDB数据库和IntegronFinder进行生物信息学分析。进行缀合分析以评估blaKPC-2通过质粒相关动员机制转移的能力。
后果
对一名住院患者进行了产KPC肺炎克雷伯菌序列型（ST）258（HA3）的高危克隆定植，该住院患者后来在住院期间被产KPC大肠杆菌ST730（HA4）感染，随后被产KPC-肺炎克雷伯菌ST11（HA15）感染。尽管属于不同的物种，但两种引起感染的菌株都具有相同的基因配置，可以在最近从布宜诺斯艾利斯自治市Alemán医院分离的其他KPC-GNB中发现的相关IncM1质粒中传播blaKPC-2。缀合分析显示，只有来自大肠杆菌HA4的pDCVEA4 KPC被成功转移，缀合频率为3.66×101。
结论
在我院一名住院患者的KPC-GNB定植和感染框架内，发现了耐多药肺炎克雷伯菌菌株ST258被ST11取代的交换。此外，blaKPC–2的基因配置在感染菌株之间的转移可能发生在医院环境中，但我们不能排除该事件发生在体内、患者体内和住院期间。
ABSTRACT
Objectives
The emergence of blaKPC-2 within nosocomial settings has become a major public health crisis worldwide. Our aim was to perform whole-genome sequencing (WGS) of three KPC-producing Gram-negative bacilli (KPC-GNB) strains isolated from a hospitalized patient to identify acquired antimicrobial resistance genes (ARGs).

Methods
WGS was performed using Illumina MiSeq-I, and de novo assembly was achieved using SPAdes. Bioinformatics analysis was done using Resfinder, AMRFinder, ISFinder, plasmidSPAdes, PlasmidFinder, MOB-suite, PLSDB database, and IntegronFinder. Conjugation assays were performed to assess the ability of blaKPC-2 to transfer via a plasmid-related mobilization mechanism.

Results
High-risk clone KPC-producing Klebsiella pneumoniae sequence type (ST) 258 (HA3) was colonizing an inpatient who later was infected by KPC-producing Escherichia coli ST730 (HA4) and subsequently by KPC-producing K. pneumoniae ST11 (HA15) during hospitalization. Although belonging to different species, both strains causing infections harbored the same gene configuration for dissemination of blaKPC-2 in related IncM1 plasmids recently found in other KPC-GNB isolated from Hospital Alemán at Ciudad Autónoma de Buenos Aires. Conjugation assays revealed that only pDCVEA4-KPC from E. coli HA4 was successfully transferred with a conjugation frequency of 3.66 × 101.

Conclusions
Interchange of multidrug-resistant K. pneumoniae lineages ST258 replaced by ST11 in the framework of colonization and infection by KPC-GNB of an inpatient from our institution was found. In addition, the transfer of the gene configuration of blaKPC–2 between infecting strains may have occurred in the nosocomial environment, but we cannot rule out that the event took place in vivo, within the patient, during hospitalization.

https://www.sciencedirect.com/science/article/pii/S2213716522002454