Ann Lab Med.  2021 Jan;41(1):86-94. 10.3343/alm.2021.41.1.86.

Antimicrobial Resistance Caused by KPC-2 Encoded by Promiscuous Plasmids of the Klebsiella pneumoniae ST307 Strain

• Lee M
• Choi TJ

• KMID: 2512737
• DOI: http://doi.org/10.3343/alm.2021.41.1.86

Abstract

Background
A lineage of Klebsiella pneumoniae that produces carbapenemase-2 (KPC-2), sequence type (ST) 307, emerged in 2017. We analyzed the complete sequences of plasmids from KPC-2-producing K. pneumoniae (KPC-Kp) ST307, investigated the antimicrobial resistance conferred by this strain, and confirmed the horizontal interspecies transmission of KPC- carbapenemase-producing Enterobacteriaceae (CPE) characteristics among Enterobacteriaceae.
Methods
We performed antimicrobial susceptibility testing, PCR analysis, multilocus sequence typing, curing tests, and whole-genome sequencing to characterize plasmid-derived KPC-2-producing Enterobacteriaceae clinical isolates.
Results
Sequence analysis of KPC-Kp strain ST307 revealed novel plasmid-located virulence factors, including a gene cluster for glycogen synthesis. Three Enterobacteriaceae strains were identified in one patient: K. pneumoniae (CPKp1825), Klebsiella aerogenes (CPEa1826), and Escherichia coli (CPEc1827). The bla KPC-2 gene from K. pneumoniae ST307 was horizontally transmitted between these strains. The plasmids could be transferred through conjugation, because all three strains of bacteria contained the type IV secretion system, pilus genes, and tra genes for conjugal transfer. The bla _KPC-2 gene was located on a truncated Tn4401 transposon. Plasmids containing the bla _KPC-2 gene could not be artificially removed; thus, the three strains could not be cured.
Conclusions
The ease of horizontal transfer of KPC-Kp ST307 carbapenem resistance has serious public health and epidemiological implications. This study provides a better understanding of the genetic characteristics that can contribute to the growth and spread of KPC-Kp ST307, and their association with antimicrobial resistance genes.

Keyword

Klebsiella pneumoniae; KPC-Kp ST307; bla _KPC gene; horizontal transfer; Tn4401; glycogen synthesis

Figure

• 

  Fig. 1 Genetic organization of plasmids associated with blaKPC. (A) Circular map of pKPBHS_25_1 with two replication origins for the IncFIB (K) and IncFII (K) groups, as well as a copper-transporting efflux system (CusCFBA) and five putative virulence clusters. (B) Variant pKPBHS_25_1 plasmids identified in ST307. (C) Circular map of pECBHS_25_2 containing ΔTn4401a with blaCTX-M-15, blaKPC-2, blaSHV-182, blaTEM-1B, and conjugative elements. Each arrow indicates plasmid scaffold genes and their direction of transcription. The locus Tra is indicated by sky blue arrows, indicating the tra genes (e.g. traG, G; traF, F; traO, O). Related genes (tnpA, tnpR, and tnpM), resistance genes, and insertion sequences are indicated by red arrows. Other genes are indicated by colored arrows as follows: orange, transcription, and translation genes; blue, CusCFBA; green, clusters encoding putative virulence determinants. In the plasmid circle, the Tn3 transposon is indicated in red, ΔTn4401a with blaKPC-2 in pink and conjugative elements in sky blue.

• 

  Fig. 2 Linear comparison of the genetic surroundings of blaKPC-2. (A) One fragment unit shows pink, yellow green, and green on the contig. Gene contents were similar along the synteny on the contig. The fragment was well preserved, but inversion existed. Most genes were well preserved. (B) pKPBHS_25_2, pEABHS_26_2, and pECBHS_27_1 are denoted by arrows and colored based on gene-functional classification. Other genes are indicated by colored arrows as follows: green, plasmid replication; blue, plasmid stability; orange, transcription and translation; sky blue, conjugative elements; pink, recombination and repair; red, antimicrobial resistance; purple, other genes.

• 

  Fig. 3 Genetic organization of plasmids pEABHS_26_2 and pECBHS_27_1. (A) Circular map of pEABHS_26_2 containing ΔTn4401a harboring blaKPC-2 and blaSHV-182 genes. (B) Circular map of pECBHS_27_1 containing ΔTn4401a harboring blaKPC-2 and conjugal transfer genes. In the plasmid circle, the Tn3 transposon is indicated in red, ΔTn4401a with blaKPC-2 in pink and conjugative elements in sky blue.

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