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

Hui Han, Haijian Zhou, Haishan Li, Yuan Gao, Zhi Lu, Kongxin Hu and Baoliang Xu
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Hui Han: Chinese Academy of Inspection and Quarantine, Beijing 100123, China
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
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

IJERPH, 2013, vol. 10, issue 7, 1-12

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 .

Keywords: Klebsiella pneumoniae; pulsed-field gel electrophoresis; molecular subtyping (search for similar items in EconPapers)
JEL-codes: I I1 I3 Q Q5 (search for similar items in EconPapers)
Date: 2013
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