Optimization of pulse-field gel electrophoresis for Bartonella subtyping.
Bartonella is a significant human pathogen and is the world's most common bacterial zoonosis acquired from companion animals. However, there is no uniform method for Pulse-Field Gel Electrophoresis (PFGE) for Bartonella population genetics studies. Further, some genes of Bartonella can mutate frequently and may affect the use of PFGE for Bartonella. Here we designed methods to solve these problems. We standardized the bacterial concentration, selected the appropriate digestion enzyme, optimized the electrophoretic parameters and characterized reproducibly two Bartonella species strains. Thus we optimized the PFGE procedure and determined how often Bartonella mutated. Our data shows a practical protocol for inter- and intra-species identification of Bartonella and was reproducible using two species strains that showed no mutation occurred after two passages for B. elizabethae; but mutation did occur in B. henselae.
Xu
Chuanbin
C
National Institute for Communicatable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, P.O. Box 5, Changping, Beijing 102206, PR China.
Liu
Qiyong
Q
Diao
Baowei
B
Kan
Biao
B
Song
Xiuping
X
Li
Dongmei
D
D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics; D015373:Bacterial Typing Techniques / Q000379:methods*; D001473:Bartonella / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D001474:Bartonella Infections / Q000382:microbiology*; D002415:Cats; D004197:Disease Reservoirs / Q000382:microbiology; D016521:Electrophoresis, Gel, Pulsed-Field / Q000379:methods*; D006801:Humans; D008969:Molecular Sequence Data; D010802:Phylogeny
D001426:Bacterial Proteins