Ann Lab Med.  2015 Nov;35(6):595-601. 10.3343/alm.2015.35.6.595.

Distribution of beta-Lactamase Genes Among Carbapenem-Resistant Klebsiella pneumoniae Strains Isolated From Patients in Turkey

• Iraz M
• Ozad Duzgun A
• Sandalli C
• Doymaz MZ
• Akkoyunlu
• Saral A
• Peleg AY
• Ozgumus OB
• Beris FS
• Karaoglu H
• Copur Cicek A

• KMID: 2363262
• DOI: http://doi.org/10.3343/alm.2015.35.6.595

Abstract

BACKGROUND
The emergence of carbapenem-resistant Klebsiella pneumoniae poses a serious problem to antibiotic management. We investigated the beta-lactamases in a group of carbapenem-resistant K. pneumoniae clinical isolates from Turkey.
METHODS
Thirty-seven strains of K. pneumoniae isolated from various clinical specimens were analyzed by antimicrobial susceptibility testing, PCR for the detection of beta-lactamase genes, DNA sequencing, and repetitive extragenic palindronic (REP)-PCR analysis.
RESULTS
All 37 isolates were resistant to ampicillin, ampicillin/sulbactam, piperacillin, piperacillin/tazobactam, ceftazidime, cefoperazone/sulbactam, cefepime, imipenem, and meropenem. The lowest resistance rates were observed for colistin (2.7%), tigecycline (11%), and amikacin (19%). According to PCR and sequencing results, 98% (36/37) of strains carried at least one carbapenemase gene, with 32 (86%) carrying OXA-48 and 7 (19%) carrying NDM-1. No other carbapenemase genes were identified. All strains carried a CTX-M-2-like beta-lactamase, and some carried SHV- (97%), TEM- (9%), and CTX-M-1-like (62%) beta-lactamases. Sequence analysis of bla(TEM) genes identified a bla(TEM-166) with an amino acid change at position 53 (Arg53Gly) from bla(TEM-1b), the first report of a mutation in this region. REP-PCR analysis revealed that there were seven different clonal groups, and temporo-spatial links were identified within these groups.
CONCLUSIONS
Combinations of beta-lactamases were found in all strains, with the most common being OXA-48, SHV, TEM, and CTX-M-type (76% of strains). We have reported, for the first time, a high prevalence of the NDM-1 (19%) carbapenemase in carbapenem-resistant K. pneumoniae from Turkey. These enzymes often co-exist with other beta-lactamases, such as TEM, SHV, and CTX-M beta-lactamases.

Keyword

bla(TEM-166); Colistin; K. pneumoniae; NDM-1; OXA-48

MeSH Terms

Anti-Bacterial Agents/*pharmacology
Bacterial Proteins/*genetics/metabolism
Carbapenems/*pharmacology
DNA, Bacterial/chemistry/genetics/metabolism
Drug Resistance, Bacterial
Genotype
Humans
Klebsiella Infections/diagnosis/microbiology
Klebsiella pneumoniae/*drug effects/enzymology/isolation & purification
Microbial Sensitivity Tests
Polymerase Chain Reaction
Sequence Analysis, DNA
Turkey
beta-Lactamases/*genetics/metabolism
Anti-Bacterial Agents
Bacterial Proteins
Carbapenems
DNA, Bacterial
beta-Lactamases

Figure

• 

  Fig. 1 Repetitive extragenic palindronic (REP)-PCR profiles of 37 Klebsiella pneumoniae isolates. Seven different genotypes were observed, and most isolates clustered into genotypes 1, 3, and 6.

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