TY - JOUR
T1 - PCR-based ORF typing of Klebsiella pneumoniae for rapid identification of global clones and transmission events
AU - Nonogaki, Rina
AU - Iijima, Anna
AU - Kawamura, Kumiko
AU - Kayama, Shizuo
AU - Sugai, Motoyuki
AU - Yagi, Tetsuya
AU - Arakawa, Yoshichika
AU - Doi, Yohei
AU - Suzuki, Masahiro
N1 - Publisher Copyright:
© 2022 The Society for Applied Microbiology.
PY - 2022/9
Y1 - 2022/9
N2 - Aims: Klebsiella pneumoniae is a major cause of healthcare-associated infections. In this study, we aimed to develop a rapid and simple genotyping method that can characterize strains causing nosocomial infections. Methods and results: The PCR-based open reading frame (ORF) typing (POT) method consists of two multiplex PCR reactions that were designed to detect 25 ORFs specific to bacterial genetic lineages, species, antimicrobial-resistant genes (blaCTX-M group-1, blaCTX-M group-9, blaIMP and blaKPC), a capsular K1-specific gene and a virulence factor gene (rmpA/A2). The electrophoresis results are then digitized. A total of 192 strains (136 clinical and 8 reference strains of K. pneumoniae, 33 clinical and 1 reference strains of K. variicola and 14 clinical strains of K. quasipneumoniae) were classified into 95, 26 and 11 POT values, respectively. The distribution patterns of ORFs among K. pneumoniae correlated well with multilocus sequence typing (MLST). Furthermore, closely related species could be distinguished and key antimicrobial resistance and hypervirulence genes were identified as part of POT. Conclusions: The POT method was developed and validated for K. pneumoniae. In comparison to MLST, the POT method is a rapid and easy genotyping method for monitoring transmission events by K. pneumoniae in clinical microbiology laboratories. Significance and Impact of the Study: The POT method supplies clear and informative molecular typing results for K. pneumoniae. The method would facilitate molecular epidemiological analysis in infection control and hospital epidemiology investigations.
AB - Aims: Klebsiella pneumoniae is a major cause of healthcare-associated infections. In this study, we aimed to develop a rapid and simple genotyping method that can characterize strains causing nosocomial infections. Methods and results: The PCR-based open reading frame (ORF) typing (POT) method consists of two multiplex PCR reactions that were designed to detect 25 ORFs specific to bacterial genetic lineages, species, antimicrobial-resistant genes (blaCTX-M group-1, blaCTX-M group-9, blaIMP and blaKPC), a capsular K1-specific gene and a virulence factor gene (rmpA/A2). The electrophoresis results are then digitized. A total of 192 strains (136 clinical and 8 reference strains of K. pneumoniae, 33 clinical and 1 reference strains of K. variicola and 14 clinical strains of K. quasipneumoniae) were classified into 95, 26 and 11 POT values, respectively. The distribution patterns of ORFs among K. pneumoniae correlated well with multilocus sequence typing (MLST). Furthermore, closely related species could be distinguished and key antimicrobial resistance and hypervirulence genes were identified as part of POT. Conclusions: The POT method was developed and validated for K. pneumoniae. In comparison to MLST, the POT method is a rapid and easy genotyping method for monitoring transmission events by K. pneumoniae in clinical microbiology laboratories. Significance and Impact of the Study: The POT method supplies clear and informative molecular typing results for K. pneumoniae. The method would facilitate molecular epidemiological analysis in infection control and hospital epidemiology investigations.
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U2 - 10.1111/jam.15701
DO - 10.1111/jam.15701
M3 - Article
C2 - 35797348
AN - SCOPUS:85134225621
SN - 1364-5072
VL - 133
SP - 2050
EP - 2062
JO - Journal of Applied Microbiology
JF - Journal of Applied Microbiology
IS - 3
ER -