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Background   Klebsiella pneumoniae  is the most frequent KPC-producing bacteria. The  bla  KPC  gene is frequently embedded in Tn4401 transposon, and less frequently in non-Tn4401 elements (NTE KPC ) variants I-III. The first case of KPC in the UC-CHRISTUS Clinical Hospital was detected in  Pseudomonas aeruginosa . Soon after this event, KPC was detected in 2 additional  Pseudomonas aeruginosa , 3  Escherichia coli , 3  Enterobacter cloacae , 3  Klebsiella pneumoniae,  and 1  Citrobacter freundii , isolated from 6 different patients. We aimed to elucidate the possible mechanisms of genetic transfer and dissemination of the  bla  KPC  gene among isolates of this multispecies outbreak. A molecular epidemiology analysis of the above mentioned clinical isolates ( n  = 13) through Multi-Locus Sequence Typing, plasmid analysis, Pulsed-Field Gel-Electrophoresis, and Whole-genome sequencing (WGS) was performed.    Results  High-risk sequence types were found:  K. pneumoniae  ST11,  P. aeruginosa  ST654, and  E. cloacae  ST114. All enterobacterial isolates were not clonal except for 3  E. coli  isolated from the same patient. WGS analysis in 6 enterobacterial isolates showed that 4 of them had  bla  KPC  embedded in a novel variant of NTE KPC  designated NTE KPC -IIe. Upstream of  bla  KPC  gene there was a 570 pb truncated  bla  TEM-1  gene followed by an insertion sequence that was 84% similar to ISEc63, a 4473 bp element of the Tn3 family. Downstream the  bla  KPC  gene there was a truncated ISKpn6 gene, and the inverted repeat right sequence of Tn4401. The ISec63-like element together with the  bla  KPC  gene plus Tn4401 remnants were inserted in the Tra operon involved in conjugative transfer of the plasmid. This NTE was carried in a broad host-range IncN plasmid.  P. aeruginosa  isolates carried  bla  KPC  gene embedded in a typical Tn4401b transposon in a different plasmid, suggesting that there was no plasmid transfer between  Enterobacteriaceae  and  P. aeruginosa  as initially hypothesized.    Conclusions  Most enterobacterial isolates had  bla  KPC  embedded in the same NTE KPC -IIe element, suggesting that this multispecies KPC outbreak was due to horizontal gene transfer rather than clonal spread. This poses a greater challenge to infection control measures often directed against containment of clonal spread.    Supplementary Information  The online version contains supplementary material available at 10.1186/s12866-021-02169-3.

A multispecies outbreak of carbapenem-resistant bacteria harboring the blaKPC gene in a non-classical transposon element