First description of NDM-1-, KPC-2-, VIM-2- and IMP-4-producing Klebsiella pneumoniae strains in a single Chinese teaching hospital

2014 ◽  
Vol 143 (2) ◽  
pp. 376-384 ◽  
Author(s):  
Y. LIU ◽  
L.-G. WAN ◽  
Q. DENG ◽  
X.-W. CAO ◽  
Y. YU ◽  
...  

SUMMARYA total of 180 non-duplicate carbapenem-resistant Klebsiella pneumoniae isolates were recovered from patients hospitalized between December 2010 and January 2012 at a Chinese hospital. Eight KPC-2, four NDM-1, one VIM-2, and five KPC-2 plus IMP-4 producers were identified and all were multidrug resistant due to the presence of other resistance determinants, including extended-spectrum β-lactamases (CTX-M-15, SHV-12), 16S rRNA methylases (armA, rmtB) and plasmid-mediated quinolone-resistance determinants (qnrA, B, S, aac(6′)-Ib-cr). Nine K. pneumoniae clones (Kpn-A1/ST395, Kpn-A3/ST11, Kpn-A2/ST134, Kpn-B/ST263, Kpn-C/ST37, Kpn-D/ST39, Kpn-E/ST1151, Kpn-F/ST890, Kpn-G/ST1153) were identified. blaKPC-2 was located on transferable ~65 kb IncL/M (ST395, ST11, ST134, ST39) and ~100 kb IncA/C (ST37, ST1153, ST890) plasmids, respectively. On the other hand, blaNDM-1 was associated with a ~70 kb IncA/C plasmid (ST263). However, non-typable plasmids of ~40 kb containing blaVIM-2 were detected in the ST1151 clone. This work reports the first co-occurrence of four diverse types of carbapenemase of K. pneumoniae clones from a single hospital in China. IncA/C, IncL/M, and other successful plasmids may be important for the dissemination of carbapenemases, producing a complex epidemiological picture.

2006 ◽  
Vol 50 (11) ◽  
pp. 3861-3866 ◽  
Author(s):  
Ying-Tsong Chen ◽  
Hung-Yu Shu ◽  
Ling-Hui Li ◽  
Tsai-Lien Liao ◽  
Keh-Ming Wu ◽  
...  

ABSTRACT A plasmid containing the qnrS quinolone resistance determinant and the gene encoding the SHV-2 β-lactamase has been discovered from a clinical Klebsiella pneumoniae strain isolated in Taiwan. The complete 98-kb sequence of this plasmid, designated pK245, was determined by using a whole-genome shotgun approach. Transfer of pK245 conferred low-level resistance to fluoroquinolones in electroporant Escherichia coli epi300. The sequence of the immediate region surrounding qnrS in pK245 is nearly identical (>99% identity) to those of pAH0376 from Shigella flexneri and pINF5 from Salmonella enterica serovar Infantis, the two other qnrS-carrying plasmids reported to date, indicating a potential common origin. Other genes conferring resistance to aminoglycosides (aacC2, strA, and strB), chloramphenicol (catA2), sulfonamides (sul2), tetracycline (tetD), and trimethoprim (dfrA14) were also detected in pK245. The dfrA14 gene is carried on a class I integron. Several features of this plasmid, including three separate regions containing putative replicons, a partitioning-control system, and a type II restriction modification system, suggest that it may be able to replicate and adapt in a variety of hosts. Although no critical conjugative genes were detected, multiple insertion sequence elements were found scattered throughout pK245, and these may facilitate the dissemination of the antimicrobial resistance determinants. We conclude that pK245 is a chimera which acquired its multiple antimicrobial resistance determinants horizontally from different sources. The identification of pK245 plasmid expands the repertoire of the coexistence of quinolone and extended-spectrum-β-lactam resistance determinants in plasmids carried by various species of the family Enterobacteriaceae in different countries.


Author(s):  
Marie N. Tuo ◽  
Augustin E. Anoh ◽  
Zéphirin O. Wayoro ◽  
Baba Coulibaly ◽  
Pacome Monemo ◽  
...  

Aims: The aims of the present study were to investigate the presence of Plasmid-Mediated Quinolone Resistance (PMQR) determinants and the association of these determinants with Extended Spectrum Beta-Lactamases (ESBLs) genes in ESBL-producing Klebsiella pneumoniae isolates from Teaching Hospital of Bouaké, Côte d’Ivoire. Study Design: It is a retrospective study. Place of Study: Bacteriology-Virology Laboratory of Teaching Hospital, Bouaké, Côte d'Ivoire. Methodology: From January 2015 to December 2016, 96 ESBL-producing Klebsiella pneumoniae isolates were collected from several specimens. Antimicrobial susceptibility of isolates was tested using the standard disk-diffusion method on Mueller-Hinton and interpretation according to recommendations of the 2017 EUCAST. These isolates analyzed for the detection of ESBL (blaCTX-M, blaTEM and blaSHV) and PMQR genes (aac(6’)-Ib-cr, qnrB and qnrS) using simplex PCR. Results: Of the 96 ESBL-producing strains, 85 (88.55%) harbored at least one of the ESBL genes tested. Out of the 85 strains encoding ESBL genes, 96.47% carried blaCTX-M and 92.94% blaSHV and blaTEM genes. Eighty nine (89.6%) of the 96 ESBL producing-isolates were resistant to ciprofloxacin and 84.4% to norfloxacin. Among the 96 strains, 80 (83.33%) were found harboring at least one PMQR gene consisting of 78 (81.3%) aac(6’)-Ib-cr, 61 (63.5%) qnrB and 15 (15.6%) qnrS. Among the PMQR-positive strains, 68.4% coharbored qnrB+acc(6’)-Ib-cr genes, 10.5% qnrB+qnrS+acc(6’)-Ib-cr and 6.6% qnrS+acc(6’)-Ib-cr. The qnrB gene was always linked to aac(6’)-Ib-cr gene. Aac(6’)-Ib-cr gene showed the highest association with three ESBL genes (87.6%), followed by qnrB gene (70.6%), then qnrS (17.7%). Conclusion: The PMQR genes were highly prevalent in ESBL-producing Klebsiella pneumoniae, primarily the aac(6’)-Ib-cr gene. The high associated was observed between ESBL and PMQR genes, notably with the aac(6’)-Ib-cr gene.


2021 ◽  
Author(s):  
Yan Wang ◽  
Guoping Cai ◽  
Jinan Zhang ◽  
Xiaogang Xu ◽  
Hongzhou Lu

Abstract BackgroundThe soaring quinolone-resistance rate of Klebsiella pneumoniae, a common pathogen in immunocompromised individuals, has seriously undermined the wide applications of antimicrobials of this class. This study aimed to investigate the emerging key contributors to quinolone-resistance in multidrug resistant K. pneumoniae (MDR-KP) isolates from a clinical setting with continuing point-source infection outbreaks in Shanghai, China. ResultsBetween January and March 2017, a total of 34 K. pneumoniae isolates, including 30 carbapenem-resistant K. pneumoniae (CRKP), were selected and characterized from a teaching hospital participating in an ongoing Bacterial Resistance Surveillance Project in Shanghai, China. Two predominant high-risk CRKP clones, ST11-wzi64 and ST15-wzi19/wzi24, caused three point-source nosocomial outbreaks in intensive care unit and/or neurosurgery department potentially by respiratory-route, promoting the co-selection and evolution of multidrug-resistant determinants. Multiple quinolone resistance-determining region (QRDR) mutations occurred in isolates of ST15 (S83F, D87A; S80I), ST11 (S83I, D87G; S80I), and ST218 (D87A; S80I). Plasmid-mediated quinolone resistance determinants, qnrS1, aac(6’)-Ib-cr, oqxAB, were detected in 32 (94.1%) isolates alone or in combination, spreading accompanied with β-lactamases (mainly, KPC-2-type carbapenemase and CTX-M-type extended-spectrum β-lactamase), 16S rRNA methylases (ArmA and RmtB), and putrescine ABC transporter permease (PotI) variants, independently of QRDR-mutations. AcrR, AcrAB transcriptional repressor, was insertion-inactivated by IS5-transposase in isolates of ST11. Thirteen ompK36 variants associated with specific ST (n=7) and wzi-allele (n=9) clustered into 10 (sub)lineages in the phylogenetic tree possibly affecting the MDR phenotype and the infection outcome of isolates. Isolates of ST11, ST15, and ST218 had frameshift disruptions in OmpK35 coupled with specific GD-insertion at position 134-135 in OmpK36, all showing distinct microevolution clusters of ompK36 genotypes. Seven quinolone-susceptible isolates kept the porin genes integral, including two each CRKPs of ST13-wzi74 (carbapenemase KPC-2 and NDM-1-coproducers) and ST65-wzi72. ConclusionsUnder selective pressures, accumulation of mutations of three types (QRDR, AcrR, OmpK36/OmpK35) and acquisition of resistance-conferring genes has been continuously contributing to quinolone-resistance in clinical MDR-KP isolates, reinforcing the importance of ongoing epidemiologic surveillance on the evolution and transmission of these isolates. Our findings provided detailed mechanistic analyses and epidemiologic implications for further infection control and antibiotic stewardship initiatives.


2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Qing Zhan ◽  
Yanlei Xu ◽  
Bingjie Wang ◽  
Jingyi Yu ◽  
Xiaofei Shen ◽  
...  

Abstract Background The rate of fluoroquinolone (FQ) resistance among carbapenem-resistant Klebsiella pneumoniae (CRKP) is high. The present study aimed to investigate the distribution of fluoroquinolone resistance determinants in clinical CRKP isolates associated with bloodstream infections (BSIs). Results A total of 149 BSI-associated clinical CRKP isolates collected from 11 Chinese teaching hospitals from 2015 to 2018 were investigated for the prevalence of fluoroquinolone resistance determinants, including plasmid-mediated quinolone resistance (PMQR) genes and spontaneous mutations in the quinolone resistance-determining regions (QRDRs) of the gyrA and parC genes. Among these 149 clinical CRKP isolates, 117 (78.5%) exhibited resistance to ciprofloxacin. The GyrA substitutions (Ser83 → IIe/Phe) and (Asp87 → Gly/Ala) were found among 112 (75.2%) of 149 isolates, while the substitution (Ser80 → IIe) of ParC was found in 111 (74.5%) of the 149 isolates. In total, 70.5% (105/149) of the CRKP isolates had at least two mutations within gyrA as well as a third mutation in parC. No mutations in the QRDRs were found in 31 ciprofloxacin susceptible CRKP isolates. Eighty-nine (56.9%) of 149 were found to carry PMQR genes including qnrS1 (43.0%), aac(6′)-Ib-cr (16.1%), qnrB4 (6.0%), qnrB2 (2.7%), and qnrB1 (1.3%). Nine isolates contained two or more PMQR genes, with one carrying four [aac(6′)-Ib-cr, qnr-S1, qnrB2, and qnrB4]. The co-existence rate of PMQR determinants and mutations in the QRDRs of gyrA and parC reached 68.5% (61/89). Seventy-four (83.1%, 74/89) PMQR-positive isolates harbored extended-spectrum beta-lactamase (ESBL)-encoding genes. Multilocus sequence typing (MLST) analysis demonstrated that the ST11 was the most prevalent STs in our study. Conclusions Mutations in the QRDRs of gyrA and parC were the key factors leading to the high prevalence of fluoroquinolone resistance among BSI-associated CRKP. The co-existence of PMQR genes and mutations in the QRDRs can increase the resistance level of CRKP to fluoroquinolones in clinical settings. ST11 CRKP isolates with identical QRDR substitution patterns were found throughout hospitals in China.


2021 ◽  
Vol 12 ◽  
Author(s):  
Qi-Sen Huang ◽  
Wenjian Liao ◽  
Zhijuan Xiong ◽  
Dan Li ◽  
Fang-Ling Du ◽  
...  

Infection caused by carbapenem-resistant hypervirulent Klebsiella pneumoniae (CR-hvKP) has become a tricky health care threat in China and KPC-2 enzyme is a main factor mediating resistance to carbapenems of K. pneumoniae. Here, we report the characterization of the genetic environment of the blaKPC-2 gene in CR-hvKP clinical isolates from South China. Forty-five non-duplicated CR-hvKP isolates collected in Jiangxi Province from 2018 to 2019 were analyzed. Each of them were multidrug-resistant due to the presence not only of blaKPC-2 gene but also of other resistance determinants, including Metallo-β-lactamases (NDM-1), extended-spectrum β-lactamases (TEM-1, CTX-M-14, SHV-1), and plasmid-mediated quinolone resistance determinants (qnrS, aac(6′)-Ib-cr). After plasmid analyses of PCR-based replicon typing (PBRT), mapping PCR, amplicon sequencing, and whole-genome sequencing (WGS) were used to analyze the genetic environment of the blaKPC-2 gene. PCR analysis of pLVPK-like plasmids, Southern Blot, and mouse lethality assay were used to characterize the virulence phenotype of K. pneumoniae. Multilocus sequence typing (MLST) analysis showed ST11 CR-hvKP was the predominant clone. In conclusion, this is the first analysis of diverse genetic structures blaKPC-2 gene in CR-hvKP isolates from south China. Both the NTEKPC-I on the IncF plasmids and pLVPK-like virulence plasmids make contributions to the formation of CR-hvKP especially ST11 which need more attention.


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