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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 which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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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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