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  Table of Contents  
Year : 2017  |  Volume : 35  |  Issue : 1  |  Page : 145-146

Emergence of ST39 and ST656 extensively drug-resistant Klebsiella pneumoniae isolates in Wenzhou, China

1 Department of Clinical Laboratory, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang Province, China
2 Department of Microbiology, Infection&Immunity Program, Biomedicine Discovery Institute, Clayton 3800, Australia
3 Department of Medical Lab Science, School of Laboratory Medicine and Life Science, Wenzhou Medical University, Wenzhou, Zhejiang Province, China

Date of Web Publication16-Mar-2017

Correspondence Address:
Tieli Zhou
Department of Clinical Laboratory, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang Province
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Source of Support: None, Conflict of Interest: None

DOI: 10.4103/ijmm.IJMM_16_381

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How to cite this article:
Liu H, Wilksch J, Li B, Du J, Cao J, Zhang X, Zhou T. Emergence of ST39 and ST656 extensively drug-resistant Klebsiella pneumoniae isolates in Wenzhou, China. Indian J Med Microbiol 2017;35:145-6

How to cite this URL:
Liu H, Wilksch J, Li B, Du J, Cao J, Zhang X, Zhou T. Emergence of ST39 and ST656 extensively drug-resistant Klebsiella pneumoniae isolates in Wenzhou, China. Indian J Med Microbiol [serial online] 2017 [cited 2020 Jul 5];35:145-6. Available from:

Dear Editor,

The emergence and dissemination of carbapenemase-producing extensively drug-resistant (XDR) Klebsiella pneumoniae has limited effective therapies and posed a tremendous challenge to public health.[1],[2]K. pneumoniae strains belonging to sequences types 11 (ST11), and ST258 are continually reported as pathogens that can cause epidemic infections in Asian and European countries.[3] Nevertheless, ST39 and ST656 had been detected only in sporadic cases so far. Here, we investigated the molecular characteristics of two ST39 and ST656 XDR K. pneumoniae strains recovered from a teaching hospital in Wenzhou, China.

Two K. pneumoniae clinical isolates (FK541 and FK2180) were isolated from the First Affiliated Hospital of Wenzhou Medical University during 2014–2015. They were collected from sputum and blood samples of two senior male patients at Intensive Care Units, respectively. The modified Hodge test (MHT) was positive for FK541, ethylenediaminetetraacetic acid (EDTA)-disk synergy test was negative. Nevertheless, the MHT and EDTA-disk synergy test were both positive for FK2180. They were proven as carbapenemase producers by utilising the phenotypic methods. Antimicrobial susceptibility testing was conducted by the agar dilution method. Two isolates were resistant to ceftriaxone, ceftazidime, cefotaxime, imipenem, meropenem, ertapenem, levofloxacin, ciprofloxacin, tobramycin, gentamicin, amikacin, chloromycetin and fosfomycin, but remained susceptible to polymyxin B and tigecycline. Furthermore, polymerase chain reaction amplification and sequencing demonstrated that two strains harboured blakpc-2, blaCTX-M-9, blaTEM-1, blaSHV, rmtB and aac(6')-Ib-cr genes simultaneously, along with single nucleotide mutations within the quinolone resistance-determining regions for strain FK541 in GyrA (Ser83Ile and Asp87Gly) and ParC (Ser80Ile), and for strain FK2180 in GyrA (Ser83Ile) and ParC (Ser80Ile). Mutations within the gyrA and parC genes could confer increased resistance to quinolones.[4] Taken together, the coexistence of multiple resistant genes in XDR K. pneumoniae isolates may contribute to high-level resistance to the majority of clinically available antimicrobial agents. Moreover, the plasmid location of the carbapenemase gene blaKPC-2 was manifested on an agarose gel following electrophoresis [Figure 1]a and Southern blot. A plasmid of ~54.2 kb in size for both strains was observed to hybridize to the blakpc-2 probe in a Southern blot [Figure 1]b, which could be transferred to Escherichia coli J53 through conjugation (transconjugants were designated as J541 and J2180).[5] Moreover, other resistance gene, such as blaCTX-M-9 for FK541, was also transferred successfully. The results of blaCTX-M-9 and rmtB probes were shown in [Figure 1]c and [Figure 1]d. Susceptibility tests revealed that both the transconjugants exhibited resistance or decreased susceptibility to cephalosporins and carbapenems, compared to E. coli J53. These data displayed the resistance determinant was transferable among species. With regard to genotypic relatedness, multilocus sequence typing revealed that two KPC-2-producing K. pneumoniae strains were typed into two rare genotypes. Strain FK541 belonged to ST39 and strain FK2180 belonged to ST656. To the best of our knowledge, this is the first description of ST656 K. pneumoniae carrying blaKPC-2 in China.
Figure 1: Plasmid analysis and southern blot hybridization of the two KPC-2-producing K. pneumoniae isolates and transconjugants. (a) Plasmid DNA of two Klebsiella pneumoniae isolates and transconjugants was separated by agarose gel electrophoresis; Escherichia coli V517 as the marker. (b) Southern blot hybridization of the gel with the blakpc probe. (c) Southern blot hybridization of the gel with the blaCTX-M-9 probe. (d) Southern blot hybridization of the gel with the rmtB probe.

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In conclusion, our study certified that KPC-2-producing K. pneumoniae strains of ST39 and ST656 are emerging in Wenzhou, the two K. pneumoniae isolates harbouring plasmids carrying the blaKPC-2 gene can be transmitted among isolates by horizontal transfer. Effective surveillance should provide guidance on the utility of antimicrobial agents to treat XDR K. pneumoniae infections.

Financial support and sponsorship

This work was supported by research grants from the National Natural Science Foundation of China (no. 81171614), the Health Department of Zhejiang Province of the People's Republic of China (no. 2011KYA106) and Zhejiang Provincial Program for the cultivation of high-level innovative health talents.

Conflicts of interest

There are no conflicts of interest.

 ~ References Top

Shi W, Li K, Ji Y, Jiang Q, Wang Y, Shi M, et al. Carbapenem and cefoxitin resistance of Klebsiella pneumoniae strains associated with porin OmpK36 loss and DHA-1 ß-lactamase production. Braz J Microbiol 2013;44:435-42.  Back to cited text no. 1
Shields RK, Nguyen MH, Potoski BA, Press EG, Chen L, Kreiswirth BN, et al. Doripenem MICs and ompK36 porin genotypes of sequence type 258, KPC-producing Klebsiella pneumoniae may predict responses to carbapenem-colistin combination therapy among patients with bacteremia. Antimicrob Agents Chemother 2015;59:1797-801.  Back to cited text no. 2
Andrade LN, Vitali L, Gaspar GG, Bellissimo-Rodrigues F, Martinez R, Darini AL. Expansion and evolution of a virulent, extensively drug-resistant (polymyxin B-resistant), QnrS1-, CTX-M-2-, and KPC-2-producing Klebsiella pneumoniae ST11 international high-risk clone. J Clin Microbiol 2014;52:2530-5.  Back to cited text no. 3
Khalil D, Becker CA, Tardy F. Alterations in the quinolone resistance-determining regions and fluoroquinolone resistance in clinical isolates and laboratory-derived mutants of Mycoplasma bovis: Not all genotypes may be equal. Appl Environ Microbiol 2015;82:1060-8.  Back to cited text no. 4
Yi H, Cho YJ, Yong D, Chun J. Genome sequence of Escherichia coli J53, a reference strain for genetic studies. J Bacteriol 2012;194:3742-3.  Back to cited text no. 5


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