Ann Lab Med.  2017 Sep;37(5):398-407. 10.3343/alm.2017.37.5.398.

Multidrug Resistance Mechanisms of Carbapenem Resistant Klebsiella pneumoniae Strains Isolated in Chongqing, China

• Yan J
• Pu S
• Jia X
• Xu X
• Yang S
• Shi J
• Sun S
• Zhang L

• KMID: 2383915
• DOI: http://doi.org/10.3343/alm.2017.37.5.398

Abstract

BACKGROUND
Carbapenem-resistant Klebsiella pneumoniae (CRKP) is considered a serious global threat. However, little is known regarding the multidrug resistance (MDR) mechanisms of CRKP. This study investigated the phenotypes and MDR mechanisms of CRKP and identified their clonal characteristics.
METHODS
PCR and sequencing were utilized to identify antibiotic resistance determinants. Integron gene cassette arrays were determined by restriction fragment length polymorphism (RFLP) analysis. Multi-locus sequence typing (MLST) and pulsed-field gel electrophoresis (PFGE) were used for epidemiological analysis. Plasmids were typed by using a PCR-based replicon typing and analyzed by conjugation and transformation assays.
RESULTS
Seventy-eight strains were identified as resistant to at least one carbapenem; these CRKP strains had a high prevalence rate (38.5%, 30/78) of carbapenemase producers. Additionally, most isolates harbored MDR genes, including Extended spectrum β-lactamases (ESBLs), AmpC, and quinolone and aminoglycoside resistance genes. Loss of porin genes was observed, and Class 1 integron was detected in 66.7% of the investigated isolates. PFGE and MLST results excluded the occurrence of clonal dissemination among these isolates.
CONCLUSIONS
A high prevalence of NDM-1 genes encoding carbapenem resistance determinants was demonstrated among the K. pneumoniae isolates. Importantly, this is the first report of bla(NDM-1) carriage in a K. pneumoniae ST1383 clone in China and of a MDR CRKP isolate co-harboring bla(NDM-1), bla(KPC-2), bla(CTX-M), bla(SHV), acc(6"²)-Ib, rmtB, qnrB, and acc(6"²)-Ib-cr.

Keyword

Carbapenem-resistant; Klebsiella pneumoniae; Carbapenemases; Extended spectrum β-lactamases; Molecular epidemiology

MeSH Terms

China*
Clone Cells
Drug Resistance, Bacterial
Drug Resistance, Microbial
Drug Resistance, Multiple*
Electrophoresis, Gel, Pulsed-Field
Genes, MDR
Integrons
Klebsiella pneumoniae*
Klebsiella*
Molecular Epidemiology
Phenotype
Plasmids
Pneumonia
Polymerase Chain Reaction
Polymorphism, Restriction Fragment Length
Prevalence
Replicon

Figure

• 

  Fig. 1 Antimicrobial resistant determinants of 78 carbapenem-resistant K. pneumoniae (CRKP) isolates. (A) Antibiotic resistance profiles of CRKP isolates: ertapenem (ETP), imipenem (IPM), meropenem (MEM), ceftriaxone (CRO), ceftazidime (CAZ), cefepime (FEP), ciprofloxacin (CIP), levofloxacin (LEV), gentamicin (GM), and tobramycin (TOB). (B) Identification of resistance determinants among the 78 CRKP isolates by PCR.

• 

  Fig. 2 Dendrogram of PFGE patterns of chromosomal DNA restriction fragments from the 78 carbapenem-resistant K. pneumoniae (CRKP) isolates. (A) A dendrogram of PFGE fingerprinting of 43 CRKP isolates. (B) The other dendrogram to show relatedness of PFGE patterns of 35 CRKP isolates. A genetic similarity index scale is shown on the left of the dendrogram. Strain number, collection data, resistance determinants, integron, gene cassettes, PFGE, and MLST types are included by each PFGE lane.Abbreviations: ESBL, extended-spectrum β-lactamase; PFGE, pulsed-field gel electrophoresis; ST, sequence type.

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