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

被引：50

作者：

Han, Hui ^{[1
]}

Zhou, Haijian ^{[2
,3
]}

Li, Haishan ^{[1
]}

Gao, Yuan ^{[2
,3
]}

Lu, Zhi ^{[4
]}

Hu, Kongxin ^{[1
]}

Xu, Baoliang ^{[1
]}

机构：

[1] Chinese Acad Inspect & Quarantine, Beijing 100123, Peoples R China

[2] Chinese Ctr Dis Control & Prevent, Natl Inst Communicable Dis Control & Prevent, Beijing 102206, Peoples R China

[3] Chinese Ctr Dis Control & Prevent, State Key Lab Infect Dis Prevent & Control, Beijing 102206, Peoples R China

[4] Gaoling Ctr Dis Control & Prevent, Xian 710200, Peoples R China

来源：

INTERNATIONAL JOURNAL OF ENVIRONMENTAL RESEARCH AND PUBLIC HEALTH | 2013年 / 10卷 / 07期

关键词：

Klebsiella pneumoniae; pulsed-field gel electrophoresis; molecular subtyping; RESISTANT STAPHYLOCOCCUS-AUREUS; LIVER-ABSCESS; ESCHERICHIA-COLI; TYPING SYSTEMS; PROTOCOL; SPREAD; ENTEROBACTERIACEAE; SURVEILLANCE; VALIDATION; STRAINS;

D O I：

10.3390/ijerph10072720

中图分类号：

X [环境科学、安全科学];

学科分类号：

08 ; 0830 ;

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.

引用

页码：2720 / 2731

页数：12

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