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<p>Carbapenemase genes play important role in the formation and dissemination of carbapenem-resistant <i>Klebsiella pneumoniae</i> (CRKP). The aim of this study was to investigate the distribution of carbapenemase genes in clinical CRKP isolates. Eighty-three clinical CRKPs were collected and antimicrobial susceptibility testing was conducted. Whole-genome sequencing was performed to analyze carbapenemase genes and genetic environments of <i>bla</i><sub>KPC-2</sub>. Recombined plasmids containing <i>bla</i><sub>KPC-2</sub> with and without IS<i>Kpn14</i> were constructed. The impact of IS<i>Kpn14</i> insertion on <i>bla</i><sub>KPC-2</sub> expression was detected by using antimicrobial susceptibility testing and reverse transcription quantitative real‑time PCR. Carbapenemase genes were detected in 77 CRKPs, mainly <i>bla</i><sub>KPC-2</sub> (70/77). ST15-KL19 (55/83) and ST11-KL64 (13/83) were dominant clones. A new sequence type, ST6816, carrying <i>bla</i><sub>NDM-1</sub> was detected in one CRKP. The genetic environments of <i>bla</i><sub>KPC-2</sub>were divided into nine types, and IS<i>26</i>-ΔTn<i>3</i>-<i>bla</i><sub>KPC-2</sub>-Tn<i>1721</i> was the dominant type (58/70). IS<i>Kpn27</i>-IS<i>Kpn14</i>-<i>bla</i><sub>KPC-2</sub>-ΔIS<i>Kpn6</i> was detected in one CRKP. With the insertion of IS<i>Kpn14</i>, the minimum inhibitory concentrations of several cephalosporins and carbapenems were significantly decreased. The relative transcription level of <i>bla</i><sub>KPC-2</sub> in plasmid containing IS<i>Kpn14</i> decreased by 14.23-fold when compared with that without IS<i>Kpn14</i>. To the best of our knowledge, IS<i>Kpn27</i>-IS<i>Kpn14</i>-<i>bla</i><sub>KPC-2</sub>-ΔIS<i>Kpn6</i> was reported for the first time in this study, and insertion of IS<i>Kpn14</i> significantly reduced the expression of <i>bla</i><sub>KPC-2</sub>.</p>

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Molecular Characterization of Carbapenemase Genes in Carbapenem-Resistant Klebsiella pneumoniae Isolates

  • 期刊论文

  • 发表日期：

    2025年7月29日

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  • Suting Li,
  • Keying Zhu,
  • Ziyan Guo,
  • Quhao Wei

摘要

Carbapenemase genes play important role in the formation and dissemination of carbapenem-resistant Klebsiella pneumoniae (CRKP). The aim of this study was to investigate the distribution of carbapenemase genes in clinical CRKP isolates. Eighty-three clinical CRKPs were collected and antimicrobial susceptibility testing was conducted. Whole-genome sequencing was performed to analyze carbapenemase genes and genetic environments of blaKPC-2. Recombined plasmids containing blaKPC-2 with and without ISKpn14 were constructed. The impact of ISKpn14 insertion on blaKPC-2 expression was detected by using antimicrobial susceptibility testing and reverse transcription quantitative real‑time PCR. Carbapenemase genes were detected in 77 CRKPs, mainly blaKPC-2 (70/77). ST15-KL19 (55/83) and ST11-KL64 (13/83) were dominant clones. A new sequence type, ST6816, carrying blaNDM-1 was detected in one CRKP. The genetic environments of blaKPC-2were divided into nine types, and IS26-ΔTn3-blaKPC-2-Tn1721 was the dominant type (58/70). ISKpn27-ISKpn14-blaKPC-2-ΔISKpn6 was detected in one CRKP. With the insertion of ISKpn14, the minimum inhibitory concentrations of several cephalosporins and carbapenems were significantly decreased. The relative transcription level of blaKPC-2 in plasmid containing ISKpn14 decreased by 14.23-fold when compared with that without ISKpn14. To the best of our knowledge, ISKpn27-ISKpn14-blaKPC-2-ΔISKpn6 was reported for the first time in this study, and insertion of ISKpn14 significantly reduced the expression of blaKPC-2.