title: Optimization of pulse-field gel electrophoresis for Borrelia burgdorferi subtyping.
abstract: OBJECTIVE
  To optimize the performance of Pulsed-Field Gel Electrophoresis (PFGE) for the comparison of inter-laboratory results and information exchange of Borrelia burgdorferi subtyping.
  
  
  METHODS
  A 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.
  
  
  RESULTS
  The 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.
  
  
  CONCLUSIONS
  PFGE can be used as a valuable test for routine genospecies identification of B. burgdorferi.
authors.affiliation: 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; Department of Infectious Disease, The First Hospital, Jilin University, Changchun 130021, Jilin, China.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Geng
authors.forename: Xue Xia
authors.initials: XX
authors.lastname: Hou
authors.forename: Qin
authors.initials: Q
authors.lastname: Hao
authors.forename: Hai Jian
authors.initials: HJ
authors.lastname: Zhou
authors.forename: Feng
authors.initials: F
authors.lastname: Wang
authors.forename: Kang Lin
authors.initials: KL
authors.lastname: Wan
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000378:metabolism; D015373:Bacterial Typing Techniques; D025065:Borrelia burgdorferi / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D004269:DNA, Bacterial / Q000378:metabolism; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D016521:Electrophoresis, Gel, Pulsed-Field; D006801:Humans; D018884:Ixodes; D051381:Rats
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; C035041:endodeoxyribonuclease MluI; C025651:CCCGGG-specific type II deoxyribonucleases; D015252:Deoxyribonucleases, Type II Site-Specific
keywords: Borrelia burgdorferi; Molecular subtyping; Pulse-Field Gel Electrophoresis