Comparison of different electrophoretic parameters of Pulse-Field Gel Electrophoresis for Vibrio cholerae subtyping.
Molecular subtyping is used to distinguish pathogenic bacterial strains during epidemiological surveys of infectious diseases and the discovery of novel pathogens. There is a need to standardize protocols involving Pulse-Field Gel Electrophoresis (PFGE), a crucial method in molecular subtyping, to make inter-laboratory results comparable and information exchange possible. The PFGE pattern varies with electrophoretic parameters (EPs), so it is important to select the parameter that can distinguish the patterns better. To optimize EPs of PFGE for Vibrio cholerae, we analyzed 24 isolates of V. cholerae O1 biotype El Tor and 26 isolates of O139 by PFGE with Not|. We used four different EPs and compared the similarity coefficients from the four groups by Friedman test. Based on the principle that the electrophoretic parameter producing smaller similarity in the PFGE patterns has higher level of discriminatory power, the one producing the smallest similarity coefficients is considered to be optimal. This method could be applied to determine the optimal molecular subtyping protocol for other bacteria.
Zhang
Jingyun
J
National Institute for Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, P. O. Box 5, Changping, Beijing 102206, PR China.
Diao
Baowei
B
Zhang
Na
N
Cui
Zhigang
Z
Zhang
Lijuan
L
Xu
Jianguo
J
Kan
Biao
B
D015373:Bacterial Typing Techniques; D004262:DNA Restriction Enzymes / Q000378:metabolism; D004269:DNA, Bacterial / Q000032:analysis; D016521:Electrophoresis, Gel, Pulsed-Field; D041241:Vibrio cholerae O1 / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification
D004269:DNA, Bacterial; D004262:DNA Restriction Enzymes