. 2018 Aug 27;56(9):e00731-18.

doi: 10.1128/JCM.00731-18. Print 2018 Sep.

Multiplex PCR Analysis for Rapid Detection of Klebsiella pneumoniae Carbapenem-Resistant (Sequence Type 258 [ST258] and ST11) and Hypervirulent (ST23, ST65, ST86, and ST375) Strains

Fangyou Yu^#^{1

2}, Jingnan Lv^#², Siqiang Niu³, Hong Du⁴, Yi-Wei Tang^{5

6}, Johann D D Pitout^{7

8}, Robert A Bonomo^{9

10

11

12

13}, Barry N Kreiswirth¹⁴, Liang Chen¹⁵

Affiliations

¹ Department of Clinical Laboratory, Shanghai Pulmonary Hospital, School of Medicine, Tongji University, Shanghai, China.
² Department of Laboratory Medicine, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, China.
³ Department of Laboratory Medicine, The First Affiliated Hospital of Chongqing Medical University, Chongqing, China.
⁴ Department of Clinical Laboratory, The Second Affiliated Hospital of Soochow University, Suzhou, Jiangsu, China.
⁵ Department of Laboratory Medicine, Memorial Sloan Kettering Cancer Center, New York, New York, USA.
⁶ Department of Pathology and Laboratory Medicine, Weill Medical College of Cornell University, New York, New York, USA.
⁷ Department of Pathology and Laboratory Medicine, University of Calgary, Calgary, Alberta, Canada.
⁸ Department of Microbiology, Immunology and Infectious Diseases, University of Calgary, Calgary, Alberta, Canada.
⁹ Case VA Center for Antimicrobial Resistance and Epidemiology (CARES), Cleveland, Ohio, USA.
¹⁰ Research Service, Louis Stokes Cleveland VA Medical Center, Cleveland, Ohio, USA.
¹¹ Department of Medicine, Pharmacology, Molecular Biology and Microbiology, Case Western Reserve University, Cleveland, Ohio, USA.
¹² Department of Pharmacology, Case Western Reserve University, Cleveland, Ohio, USA.
¹³ Department of Molecular Biology and Microbiology, Case Western Reserve University, Cleveland, Ohio, USA.
¹⁴ Public Health Research Institute Tuberculosis Center, New Jersey Medical School, Rutgers University, Newark, New Jersey, USA.
¹⁵ Public Health Research Institute Tuberculosis Center, New Jersey Medical School, Rutgers University, Newark, New Jersey, USA Chen11@njms.rutgers.edu.

^# Contributed equally.

PMID: 29925644
PMCID: PMC6113471
DOI: 10.1128/JCM.00731-18

Multiplex PCR Analysis for Rapid Detection of Klebsiella pneumoniae Carbapenem-Resistant (Sequence Type 258 [ST258] and ST11) and Hypervirulent (ST23, ST65, ST86, and ST375) Strains

Fangyou Yu et al. J Clin Microbiol. 2018.

. 2018 Aug 27;56(9):e00731-18.

doi: 10.1128/JCM.00731-18. Print 2018 Sep.

Authors

Fangyou Yu^#^{1

2}, Jingnan Lv^#², Siqiang Niu³, Hong Du⁴, Yi-Wei Tang^{5

6}, Johann D D Pitout^{7

8}, Robert A Bonomo^{9

10

11

12

13}, Barry N Kreiswirth¹⁴, Liang Chen¹⁵

Affiliations

¹ Department of Clinical Laboratory, Shanghai Pulmonary Hospital, School of Medicine, Tongji University, Shanghai, China.
² Department of Laboratory Medicine, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, China.
³ Department of Laboratory Medicine, The First Affiliated Hospital of Chongqing Medical University, Chongqing, China.
⁴ Department of Clinical Laboratory, The Second Affiliated Hospital of Soochow University, Suzhou, Jiangsu, China.
⁵ Department of Laboratory Medicine, Memorial Sloan Kettering Cancer Center, New York, New York, USA.
⁶ Department of Pathology and Laboratory Medicine, Weill Medical College of Cornell University, New York, New York, USA.
⁷ Department of Pathology and Laboratory Medicine, University of Calgary, Calgary, Alberta, Canada.
⁸ Department of Microbiology, Immunology and Infectious Diseases, University of Calgary, Calgary, Alberta, Canada.
⁹ Case VA Center for Antimicrobial Resistance and Epidemiology (CARES), Cleveland, Ohio, USA.
¹⁰ Research Service, Louis Stokes Cleveland VA Medical Center, Cleveland, Ohio, USA.
¹¹ Department of Medicine, Pharmacology, Molecular Biology and Microbiology, Case Western Reserve University, Cleveland, Ohio, USA.
¹² Department of Pharmacology, Case Western Reserve University, Cleveland, Ohio, USA.
¹³ Department of Molecular Biology and Microbiology, Case Western Reserve University, Cleveland, Ohio, USA.
¹⁴ Public Health Research Institute Tuberculosis Center, New Jersey Medical School, Rutgers University, Newark, New Jersey, USA.
¹⁵ Public Health Research Institute Tuberculosis Center, New Jersey Medical School, Rutgers University, Newark, New Jersey, USA Chen11@njms.rutgers.edu.

^# Contributed equally.

PMID: 29925644
PMCID: PMC6113471
DOI: 10.1128/JCM.00731-18

Abstract

Carbapenem-resistant and hypervirulent Klebsiella pneumoniae strains have emerged recently. These strains are both hypervirulent and multidrug resistant and may also be highly transmissible and able to cause severe infections in both the hospital and the community. Clinical and public health needs require a rapid and comprehensive molecular detection assay to identify and track the spread of these strains and provide timely infection control information. Here, we develop a rapid multiplex PCR assay capable of distinguishing K. pneumoniae carbapenem-resistant isolates of sequence type 258 (ST258) and ST11, and hypervirulent ST23, ST65/ST375, and ST86 clones, as well as capsular types K1, K2, K locus type 47 (KL47), and KL64, and virulence genes rmpA, rmpA2, iutA, and iroN The assay demonstrated 100% concordance with 118 previously genotyped K. pneumoniae isolates and revealed different populations of carbapenem-resistant and hypervirulent strains in two collections in China and the United States. The results showed that carbapenem-resistant and hypervirulent K. pneumoniae strains are still rare in the United States, whereas in China, ∼50% of carbapenem-resistant strains carry rmpA/rmpA2 and iutA virulence genes, which are largely associated with the epidemic ST11 strains. Similarly, a high prevalence of hypervirulent strains was found in carbapenem-susceptible isolates in two Chinese hospitals, but these primarily belong to ST23, ST65/ST375, and ST86, which are distinct from the carbapenem-resistant strains. Taken together, our results demonstrated that this PCR assay can be a useful tool for molecular surveillance of carbapenem-resistant and hypervirulent K. pneumoniae strains.

Keywords: capsular polysaccharide; carbapenem resistance; hypervirulence; multiplex PCR; plasmid.

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Figures

**FIG 1**
Agarose gel electrophoresis of multiplex PCR products.

See this image and copyright information in PMC

References

1. Lee CR, Lee JH, Park KS, Jeon JH, Kim YB, Cha CJ, Jeong BC, Lee SH. 2017. Antimicrobial resistance of hypervirulent Klebsiella pneumoniae: epidemiology, hypervirulence-associated determinants, and resistance mechanisms. Front Cell Infect Microbiol 7:483. doi:10.3389/fcimb.2017.00483. - DOI - PMC - PubMed
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Multiplex PCR Analysis for Rapid Detection of Klebsiella pneumoniae Carbapenem-Resistant (Sequence Type 258 [ST258] and ST11) and Hypervirulent (ST23, ST65, ST86, and ST375) Strains

Affiliations

Multiplex PCR Analysis for Rapid Detection of Klebsiella pneumoniae Carbapenem-Resistant (Sequence Type 258 [ST258] and ST11) and Hypervirulent (ST23, ST65, ST86, and ST375) Strains

Authors

Affiliations

Abstract

Figures

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LinkOut - more resources

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