Rapid pulsed-field gel electrophoresis protocol for subtyping of Campylobacter jejuni

We developed a rapid pulsed field gel electrophoresis (PFGE) protocol for s ubtyping Campylobacter isolates based on the standardized protocols used by PulseNet laboratories for the subtyping of other food-borne bacterial path ogens. Various combinations of buffers, reagents, reaction conditions (e.g. , cell suspension concentration, lysis time, lysis temperature, and restric tion enzyme concentration), and electrophoretic parameters were evaluated i n an effort to devise a protocol that is simple, rapid, and robust. PFGE an alysis of Campylobacter isolates can be completed in 24 to 30 h using this protocol, whereas the most widely used current protocols require 3 to 4 day s to complete. Comparison of PFGE patterns obtained in six laboratories sho wed that subtyping results obtained using this protocol are highly reproduc ible.