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Int. J. Environ. Res. Public Health 2013, 10(7), 2720-2731; doi:10.3390/ijerph10072720
Article

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

1,†
,
2,†,* , 1
,
2
,
3
,
1
 and
1,*
1 Chinese Academy of Inspection and Quarantine, Beijing 100123, China 2 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 3 Gaoling Center for Diseases Control and Prevention, Xi'an 710200, China These authors contributed equally to this work.
* Authors to whom correspondence should be addressed.
Received: 9 May 2013 / Revised: 5 June 2013 / Accepted: 13 June 2013 / Published: 1 July 2013
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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 XbaI-PFGE, 11 isolates were analyzed with XbaI 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 XbaI PFGE of K. pneumoniae. By software analysis and pilot study, AvrII was chosen as another PFGE enzyme. Both XbaI- and AvrII-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 XbaI- and AvrII-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 Klebsiella pneumoniae; pulsed-field gel electrophoresis; molecular subtyping
This is an open access article distributed under the Creative Commons Attribution License (CC BY 3.0).
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Han, H.; Zhou, H.; Li, H.; Gao, Y.; Lu, Z.; Hu, K.; Xu, B. Optimization of Pulse-Field Gel Electrophoresis for Subtyping of Klebsiella pneumoniae. Int. J. Environ. Res. Public Health 2013, 10, 2720-2731.

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