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Journal of Clinical Microbiology, May 2001, p. 1889-1894, Vol. 39, No. 5
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.5.1889-1894.2001

Rapid Pulsed-Field Gel Electrophoresis Protocol for Subtyping of Campylobacter jejuni

Efrain M. Ribot,* Collette Fitzgerald, Kristy Kubota, Bala Swaminathan, and Timothy J. Barrett

Centers for Disease Control and Prevention, Atlanta, Georgia

Received 19 October 2000/Returned for modification 30 December 2000/Accepted 4 March 2001

We developed a rapid pulsed-field gel electrophoresis (PFGE) protocol for subtyping Campylobacter isolates based on the standardized protocols used by PulseNet laboratories for the subtyping of other food-borne bacterial pathogens. Various combinations of buffers, reagents, reaction conditions (e.g., cell suspension concentration, lysis time, lysis temperature, and restriction enzyme concentration), and electrophoretic parameters were evaluated in an effort to devise a protocol that is simple, rapid, and robust. PFGE analysis 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 days to complete. Comparison of PFGE patterns obtained in six laboratories showed that subtyping results obtained using this protocol are highly reproducible.

* Corresponding author. Mailing address: Centers for Disease Control and Prevention, 1600 Clifton Road, Mail Stop C03, Atlanta, GA 30333. Phone: (404) 639-3764. Fax: (404) 639-3333. E-mail: eyr4{at}cdc.gov.

Journal of Clinical Microbiology, May 2001, p. 1889-1894, Vol. 39, No. 5
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.5.1889-1894.2001


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