title: Optimization of pulse-field gel electrophoresis for subtyping of Klebsiella pneumoniae.
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.
authors.affiliation: Chinese Academy of Inspection and Quarantine, Beijing 100123, China. hanhui2002@163.com
authors.forename: Hui
authors.initials: H
authors.lastname: Han
authors.forename: Haijian
authors.initials: H
authors.lastname: Zhou
authors.forename: Haishan
authors.initials: H
authors.lastname: Li
authors.forename: Yuan
authors.initials: Y
authors.lastname: Gao
authors.forename: Zhi
authors.initials: Z
authors.lastname: Lu
authors.forename: Kongxin
authors.initials: K
authors.lastname: Hu
authors.forename: Baoliang
authors.initials: B
authors.lastname: Xu
mesh_terms: D015373:Bacterial Typing Techniques / Q000379:methods*; D004269:DNA, Bacterial / Q000235:genetics / Q000378:metabolism; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D016521:Electrophoresis, Gel, Pulsed-Field / Q000379:methods*; D007711:Klebsiella pneumoniae / Q000145:classification* / Q000235:genetics
chemical_list: D004269:DNA, Bacterial; C025580:endodeoxyribonuclease XBAI; C064053:CCTAGG-specific type II deoxyribonucleases; D015252:Deoxyribonucleases, Type II Site-Specific