Optimization of pulse-field gel electrophoresis for Borrelia burgdorferi subtyping.

Zhen Geng; Xue Xia Hou; Qin Hao; Hai Jian Zhou; Feng Wang; Kang Lin Wan

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Optimization of pulse-field gel electrophoresis for Borrelia burgdorferi subtyping.

Author: Zhen GENG ^{1
,

2} ; Xue Xia HOU ; Qin HAO ; Hai Jian ZHOU ; Feng WANG ; Kang Lin WAN
Author Information

1. State Key Laboratory for Communicable Disease Prevention and Control, National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China
2. Department of Infectious Disease, The First Hospital, Jilin University, Changchun 130021, Jilin, China.
Publication Type:Journal Article
Keywords: Borrelia burgdorferi; Molecular subtyping; Pulse-Field Gel Electrophoresis
MeSH: Animals; Bacterial Proteins; metabolism; Bacterial Typing Techniques; Borrelia burgdorferi; classification; genetics; isolation & purification; DNA, Bacterial; metabolism; Deoxyribonucleases, Type II Site-Specific; metabolism; Electrophoresis, Gel, Pulsed-Field; Humans; Ixodes; Rats
From: Biomedical and Environmental Sciences 2013;26(7):584-591
CountryChina
Language:English
Abstract:
OBJECTIVETo optimize the performance of Pulsed-Field Gel Electrophoresis (PFGE) for the comparison of inter-laboratory results and information exchange of Borrelia burgdorferi subtyping.
METHODSA panel of 34 strains of B. burgdorferi were used to optimize PFGE for subtyping. In order to optimize the electrophoretic parameters (EPs), all 34 strains of B. burgdorferi were analyzed using four EPs, yielding different Simpson diversity index (D) values and the epidemiological concordance was also evaluated.
RESULTSThe EP of a switch time of 1 s to 25 s for 13 h and 1 s to 10 s for 6 h produced the highest D value and was declared to be optimal for MluI and SmaI PFGE of B. burgdorferi. MluI and SmaI were selected as the first and second restriction enzymes for PFGE subtyping of B. burgdorferi according to discrimination and consistency with epidemiological data.
CONCLUSIONPFGE can be used as a valuable test for routine genospecies identification of B. burgdorferi.