Molecular characterization of clinical IMP-producing Klebsiella pneumoniae isolates from a Chinese Tertiary Hospital

Lai, Kaisheng; Ma, Yue; Guo, Ling; An, Jingna; Ye, Liyan; Yang, Jiyong

doi:10.1186/s12941-017-0218-9

Cited by 18 publications

(17 citation statements)

References 29 publications

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“…A 1 kb DNA ladder and lambda DNA/HindIII (Promega, Madison,WI USA) were used as DNA markers and plasmid extraction also was performed by using plasmid extraction kit (Qiagen), this experiment was repeated 3 times to confirm the results. The plasmid linearization and confirmation of the plasmid sizes were performed by S1 nuclease PFGE and lambda DNA; low-range DNA markers and E. coli V517 strain were used as standard reference [25]. PCR detection of selected βlactamase genes (bla SHV , bla TEM , bla CTXM-1 group, and bla CTXM-9 group) and colistin resistance gene (mcr-1) was performed using extracted plasmid DNA.…”

Section: Methodsmentioning

confidence: 99%

“…PCR detection of selected βlactamase genes (bla SHV , bla TEM , bla CTXM-1 group, and bla CTXM-9 group) and colistin resistance gene (mcr-1) was performed using extracted plasmid DNA. The PCR products were sequenced to confirm their identity [25,26].…”

Section: Methodsmentioning

confidence: 99%

“…Transfer of ESBL and mcr-1 genes by conjugation was performed in Luria-Bertani broth using nalidixic acidresistant E. coli DH5α as the recipient strain and conjugation experiment was repeated 5 times to confirm the results. Transconjugants were selected on LB agar supplemented with nalidixic acid (100 mg/mL) and cefotaxime (2 mg/L) or colistin (1 mg/L) (Sigma Aldrich) [25,26]. PCR detection of selected β-lactamase genes (bla SHV , bla TEM , bla CTXM-1 group, bla CTXM-9 group, and mcr-1) was performed using plasmid DNA extracted from transconjugants.…”

Section: Methodsmentioning

confidence: 99%

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Molecular characterization of extended-spectrum β-lactamase-producing Klebsiella pneumoniae from a Malaysian hospital

et al. 2019

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Multidrug-resistant (MDR) and extended-spectrum β-lactamase (ESBL)-producing Klebsiella pneumoniae associated with nosocomial infections have caused serious problems in antibiotic management with limited therapeutic choices. This study aimed to determine the genotypic and phenotypic characteristics of K. pneumoniae strains isolated from a tertiary hospital in Malaysia. Ninety-seven clinical K. pneumoniae strains were analyzed for antimicrobial susceptibility, all of which were sensitive to amikacin and colistin (except one strain), while 31.9 % and 27.8 % were MDR and ESBL producers, respectively. PCR and DNA sequencing of the amplicons indicated that the majority of MDR strains (26/27) were positive for bla TEM, followed by bla SHV (24/27), bla CTX-M-1 group (23/27), bla CTX-M-9 group (2/27), and mcr-1 (1/27). Thirty-seven strains were hypervirulent and PCR detection of virulence genes showed 38.1 %, 22.7 %, and 16.5 % of the strains were positive for K1, wabG, and uge genes, respectively. Genotyping by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) showed that these strains were genetically diverse and heterogeneous. Sequence types, ST23, ST22, and ST412 were the predominant genotypes. This is the first report of colistin-resistant K. pneumoniae among clinical strains associated with mcr-1 plasmid in Malaysia. The findings in this study have contributed to the effort in combating the increase in antimicrobial resistance by providing better understanding of genotypic characteristics and resistance mechanisms of the organisms.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Molecular characterization of extended-spectrum β-lactamase-producing Klebsiella pneumoniae from a Malaysian hospital

et al. 2019

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“…In this study, bla IMP-4 was located on an IncN type plasmid (~50 kb), and plasmids belonging to the IncN incompatible group typically have a broad host range and high transmission efficiency, and they play an important role in the transmission of clinically important resistance determinants. 23,33,34 Location of the major carbapenem resistance genes such as bla IMP , 35,36 bla KPC , 37,38 bla NDM 39,40 and bla VIM 34,41 has been found on different IncN-type plasmids. The IncN plasmids can be further divided into three subgroups, namely IncN1to IncN3, with their reference plasmids R46 (accession number AY046276), p271A 42 and pN-Cit, 43 respectively.…”

Section: Discussionmentioning

confidence: 99%

<p>Complete-Genome Sequencing and Comparative Genomic Characterization of an IMP-4 Producing <em>Citrobacter freundii</em> Isolate from Patient with Diarrhea</p>

Chi¹,

Guo²,

Zhou³

et al. 2020

IDR

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Background: Citrobacter freundii is the most common class of pathogens in the genus Citrobacter and is an important pathogen associated with certain underlying diseases or immune dysfunction. The aim of this study was to elucidate the resistance mechanism of clinically derived carbapenem-resistant C. freundii isolate and to characterize the genetic environment and delivery pattern of the IncN1 plasmid carrying the bla IMP-4 gene from C. freundii isolate. Materials and Methods: We identified a clinical isolate of C. freundii L91 carrying bla IMP-4 and performed phylogenetic analysis by whole-genome sequencing. The complete genomic sequence of L91 was obtained using the Illumina HiSeq 4000-PE150 and PacBio RS II platforms. Antimicrobial susceptibility testing was determined by the VITEK 2 system. Plasmid characteristics were presented by S1-pulsed-field gel electrophoresis (PFGE), Southern blotting and conjugation experiments. Results: S1-PFGE, Southern blot and conjugation assay confirmed the presence of bla IMP-4 genes on a conjugative plasmid in this isolate. C. freundii L91 and transconjugant L91-E. coli 600 strains both showed resistance to carbapenems. In silico analysis further showed that pIMP-4-L91 is an IncN1 plasmid with a length of 51,042 bp. Furthermore, bla IMP-4 gene was found encoded in the bla IMP-4-qacG2-aacA4-catB3 cassette array within a class 1 integron. A conserved structure sequence (ΔISKpn27-bla IMP-4-ΔISSen2-hp-hp-IS6100) was found in the upstream and downstream of the bla IMP-4. Conclusion: We performed a comprehensive phylogenetic analysis of carbapenemaseresistant C. freundii and elucidated the resistance mechanism of clinically derived C. freundii L91. Not only that, we also found that the bla IMP-4 gene is located on the IncN1 plasmid and has a horizontal transfer function and a certain ability to spread. To lower the risk of the dissemination of such C. freundii isolates in clinical settings, more surveillance is needed in the future.

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“…IMP carbapenemase, a member of the metallo-␤-lactamases capable of hydrolyzing almost all ␤-lactams, was first described in Japan in the 1990s (11). In 2011, our group first identified a novel IMP-38 variant in the neonatology department in our hospital (12).…”

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confidence: 97%

IMP-38-Producing High-Risk Sequence Type 307 Klebsiella pneumoniae Strains from a Neonatal Unit in China

Wang

Zhao

Liu

et al. 2020

mSphere

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An emerging multidrug-resistant Klebsiella pneumoniae high-risk clone of sequence type 307 (ST307) has been increasingly reported worldwide. Here, we described the genomic characteristics of an IMP-38-producing ST307 K. pneumoniae strain and investigated the prevalence of blaIMP-38 among carbapenem-resistant Klebsiella pneumoniae isolates from a tertiary care hospital in central China. A total of 14 IMP-38-producing ST307 K. pneumoniae strains were identified from 2013 to 2016, with 13 strains isolated from patients with neonatal sepsis in the neonatal ward. PacBio and Illumina whole-genome sequencing analysis performed on a representative IMP-38-producing K. pneumoniae strain, WCGKP294, showed that it contained a circular chromosome and two plasmids. Carbapenemase gene blaIMP-38 is colocated with blaCTX-M-3 in transposon Tn6382 on an IncHI5 plasmid (pWCGKP294-2). WCGKP294 harbors another IncFIB plasmid, pWCGKP294-1, carrying three copies of tandem-repeated IS26-blaSHV-2A-deoR-ygbJ-ygbK-fucA-IS26 composite transposon elements. Phylogenetic analysis placed WCGKP294 in the global ST307 cluster, distant from the U.S. (Texas) and South Africa clusters. Nevertheless, WCGKP294 does not contain the chromosomal fluoroquinolone resistance-associated mutations and IncFIIK/IncFIBK plasmid-associated blaCTX-M-15 gene that are frequently found in other global ST307 strains. IMPORTANCE We described the genome and resistome characterization of a carbapenem-resistant Klebsiella pneumoniae ST307 strain carrying blaIMP-38 in China. This report highlights that the high-risk ST307 clone continues to acquire different antimicrobial resistance genes, posing significant challenges to clinical practice, and should be closely monitored.

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Molecular characterization of clinical IMP-producing Klebsiella pneumoniae isolates from a Chinese Tertiary Hospital

Cited by 18 publications

References 29 publications

Molecular characterization of extended-spectrum β-lactamase-producing Klebsiella pneumoniae from a Malaysian hospital

Molecular characterization of extended-spectrum β-lactamase-producing Klebsiella pneumoniae from a Malaysian hospital

<p>Complete-Genome Sequencing and Comparative Genomic Characterization of an IMP-4 Producing <em>Citrobacter freundii</em> Isolate from Patient with Diarrhea</p>

IMP-38-Producing High-Risk Sequence Type 307 Klebsiella pneumoniae Strains from a Neonatal Unit in China

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