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J Clin Microbiol. 2018 Aug 27;56(9). pii: 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.

Yu F#1,2, Lv J#2, Niu S3, Du H4, Tang YW5,6, Pitout JDD7,8, Bonomo RA9,10,11,12,13, Kreiswirth BN14, Chen L15.

Author information

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

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

PMID:
29925644
PMCID:
PMC6113471
DOI:
10.1128/JCM.00731-18
[Indexed for MEDLINE]
Free PMC Article

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