Coexistence of two blaKPC-2 genes in a blaNDM-1-carrying multidrug-resistant ST15 Klebsiella pneumoniae isolate recovered from cerebrospinal fluid in China

Objectives: Carbapenem-resistant Klebsiella pneumoniae (CRKP) is increasingly reported worldwide and has posed a serious challenge for public health. Here we report the complete genome sequence of a multidrug-resistant (MDR) K. pneumoniae carrying one bla(NDM-1) and two copies of bla(KPC-2) genes isolated from a cerebrospinal fluid specimen in China. Methods: The minimal inhibitory concentrations (MICs) of 26 antimicrobial agents against K. pneumoniae strain KP46 were measured. The complete genome sequence of KP46 was determined using Illumina and Nanopore platforms. The derived short and long reads were assembled using Unicycler. Multilocus sequence typing (MLST), antimicrobial resistance genes, virulence genes, and plasmid replicons were pre-dicted in silico using the BacWGSTdb server. The phylogenetic relationship between KP46 and 454 ST15 K. pneumoniae strains obtained from NCBI GenBank database was analysed based on a core genome MLST (cgMLST) strategy. Results: K. pneumoniae strain KP46 was resistant to all antimicrobial agents tested, except for tigecycline, colistin, cefiderocol, and fosfomycin. The genome sequence of KP46 belonged to sequence type 15 (ST15), which contained seven circularized contigs comprising 5 674 521 bp, including one chromosome and six plasmids. Serval antimicrobial resistance genes were identified, including a bla(NDM-1) gene located in a 53 096 bp IncX3 plasmid, and two copies of bla(KPC-2) gene located both in a 103 807 bp IncX6 and an 88 164 bp IncFII plasmid, respectively. The most closely related strain was another ST15 strain also isolated from China with five cgMLST loci differences. Conclusion: We reported the first complete genome sequence of a K. pneumoniae ST15 clinical isolate coharbouring bla(NDM-1) and two copies of bla(KPC-2) in China. This study will provide insight into the an-timicrobial resistance mechanisms and phylogeny of carbapenem-resistant ST15 K. pneumoniae. (C) 2022 The Authors. Published by Elsevier Ltd on behalf of International Society for Antimicrobial Chemotherapy.