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Optimization of Pulse-Field Gel Electrophoresis for Subtyping of Klebsiella pneumoniae

Author

Listed:
  • Hui Han

    (Chinese Academy of Inspection and Quarantine, Beijing 100123, China
    These authors contributed equally to this work.)

  • Haijian Zhou

    (National Institute for Communicable Disease Control and Prevention, and State Key Laboratory for Infectious Disease Prevention and Control, Chinese Center for Disease Control and Prevention, Beijing 102206, China
    These authors contributed equally to this work.)

  • Haishan Li

    (Chinese Academy of Inspection and Quarantine, Beijing 100123, China)

  • Yuan Gao

    (National Institute for Communicable Disease Control and Prevention, and State Key Laboratory for Infectious Disease Prevention and Control, Chinese Center for Disease Control and Prevention, Beijing 102206, China)

  • Zhi Lu

    (Gaoling Center for Diseases Control and Prevention, Xi'an 710200, China)

  • Kongxin Hu

    (Chinese Academy of Inspection and Quarantine, Beijing 100123, China)

  • Baoliang Xu

    (Chinese Academy of Inspection and Quarantine, Beijing 100123, China)

Abstract

A total of 110 strains of Klebsiella pneumoniae were used to optimize pulsed-field gel electrophoresis (PFGE) for subtyping of K. pneumoniae . For optimization of electrophoresis parameters (EPs) of Xba I-PFGE, 11 isolates were analyzed with Xba I digestion using three EPs. The EP of a switch time of 6 to 36 s for 18.5 h gave clearest patterns and was declared the optimal EP for Xba I PFGE of K. pneumoniae . By software analysis and pilot study, Avr II was chosen as another PFGE enzyme. Both Xba I- and Avr II-PFGE gave D -values higher than 0.99 for 69 K. pneumoniae isolated from different sources . Our results also showed good typeability, reproducibility of both Xba I- and Avr II-PFGE for K. pneumoniae subtyping. Furthermore, the established PFGE method also had good discriminatory power to distinguish outbreak K. pneumoniae strains and a high degree of consistency with multilocus sequence typing method. A rapid PFGE protocol was established here, which could be used for genotyping and other researches of K. pneumoniae .

Suggested Citation

  • Hui Han & Haijian Zhou & Haishan Li & Yuan Gao & Zhi Lu & Kongxin Hu & Baoliang Xu, 2013. "Optimization of Pulse-Field Gel Electrophoresis for Subtyping of Klebsiella pneumoniae," IJERPH, MDPI, vol. 10(7), pages 1-12, July.
    • Handle: RePEc:gam:jijerp:v:10:y:2013:i:7:p:2720-2731:d:26819
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