Pulsed field gel electrophoresis for determining the molecular homology of Campylobacter jejuni isolates

Abstract:

Abstract: [Objective] To establish a genetic typing map for Campylobacter jejuni in China by using Pulsed Field Gel Electrophoresis (PFGE). [Methods] A PFGE protocol was developed for subtyping C. jejuni isolates from different sources. The effect of cell suspension concentration, Seakem Gold? agarose gel strength, proteinase K density, washing style, and restriction enzyme concentration were also evaluated. [Results] DNA from 37 isolated strains of C. jejuni digested with restriction enzyme Sma produced PFGE profiles that distinguished these strains to the level of species, and resulted in 6 to 24 bands of electrophoresis. A phyletic evolution tree showed that the isolates could be separated into four genetic lineages, and possessed dominance among lineage Ⅳ. The distribution of isolated strains from different sources overlapped each other in all groups. [Conclusion] PFGE can be used for the investigation of molecular epidemiological patterns of C. jejuni and for tracing the source of C. jejuni in clinical patients. Preliminary data showed that human Campylobacteriosis was related to C. jejuni of animal origin, particularly from the chicken.