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The emergence of carbapenem-resistant  Enterobacteriaceae  poses a significant threat to public health worldwide. Here, we reported a multidrug-resistant  Escherichia coli  strain with two different  bla  NDM–  5 -carrying plasmids from China. Illumina short-read and MinION long-read whole genome sequencing were performed. Genomic analysis found that one  bla  NDM–  5  gene together with  mphA  was located on a 55-kb IncX3 plasmid, while the other  bla  NDM–  5  gene was on a novel 68-kb IncFII plasmid. Susceptibility testing and quantitative reverse transcription PCR results further indicated that the transconjugants with the IncX3 plasmid exhibited higher-level carbapenem resistance and expression of  bla  NDM–  5  than those with both plasmids or the IncFII plasmid. Two other β-lactamase genes ( bla   CTX–M–   15  and  bla  OXA–  1 ) were also detected on another 160-kb IncF plasmid. This is the first report of coexistence of two  bla  NDM–  5 -carrying plasmids in a single bacterial isolate, highlighting the genetic complexity of NDM-5 carbapenemase circulation, and the urgent need for continued active surveillance.

Coexistence of Two blaNDM–5 Genes Carried on IncX3 and IncFII Plasmids in an Escherichia coli Isolate Revealed by Illumina and Nanopore Sequencing