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

International Journal of Environmental Research and Public Health

Hui Han²  Haijian Zhou¹  Haishan Li²  Yuan Gao¹  Zhi Lu³  Kongxin Hu² 

[1] 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; E-Mail:;Chinese Academy of Inspection and Quarantine, Beijing 100123, China; E-Mails:;Gaoling Center for Diseases Control and Prevention, Xi’an 710200, China; E-Mail:

关键词: Klebsiella pneumoniae;    pulsed-field gel electrophoresis;    molecular subtyping;   

DOI  :  10.3390/ijerph10072720

来源: mdpi

【 摘 要 】

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.

【 授权许可】

CC BY   
© 2013 by the authors; licensee MDPI, Basel, Switzerland.