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Three carbapenem-resistant  Klebsiella pneumoniae  (CRKP; strains KP-426, KP-C76, and KP-CT77) were isolated from a patient with severe burns during the treatment of colistin and tigecycline. Single-nucleotide polymorphism typing showed that three ST11 CRKP were clonally related. Three isolates harbored the same set of antimicrobial resistance genes.  bla  KPC-2 ,  bla  SHV-12 ,  bla  TEM-1 , and  rmtB  genes were located on the same 128,928-bp IncFII/IncR plasmid.  Tet (A),  catA2 ,  sul2 , and  dfrA14  genes were located on a plasmid with an unknown Inc-type.  bla  SHV-11 ,  fosA , and  aadA2  were chromosomal genes. An IS 1  and an IS Kpn14  were found in the promoter region of the  mgrB  gene of two colistin-resistant CRKP,  K. pneumoniae  KP-C76, and KP-CT77, respectively. A novel amino acid substitution, G300E, was identified in the type 1 Tet(A) variant of  K. pneumoniae  KP-CT77 which exhibited high-level tigecycline resistance compared to strains KP-426 and KP-C76 (MIC of 32, 4, and 4mg/l, respectively). Conjugation and cloning experiments confirmed that the mutated Tet(A) resulted in a 4-fold increase in tigecycline minimal inhibitory concentration (MIC) of  Escherichia coli . Three CRKP belonged to the K64 serotype and possessed a similar IncHI1B/repB virulence plasmid carrying  rmpA ,  rmpA2 , and  iucABCDiutA . The survival rates of  Galleria Mellonella  injected with  K. pneumoniae  KP-426, KP-C76, and KP-CT77 were 4.2, 20.8, and 8.3%, respectively. The emergence of colistin and tigecycline resistance in carbapenem-resistant hypervirulent  K. pneumoniae  posed a serious threat to clinical anti-infective therapy. The type 1 Tet(A) variant carrying G300E mutation, which conferred significantly elevated tigecycline MIC and was located on a conjugative plasmid, needs attention.

In vivo Emergence of Colistin and Tigecycline Resistance in Carbapenem-Resistant Hypervirulent Klebsiella pneumoniae During Antibiotics Treatment