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Journal of Clinical Microbiology, January 2003, p. 27-33, Vol. 41, No. 1
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.1.27-33.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Subtyping of Salmonella enterica Serotype Enteritidis Strains by Manual and Automated PstI-SphI Ribotyping

Clifford G. Clark,1* Tamara M. A. C. Kruk,1 Louis Bryden,1 Yolanda Hirvi,2 Rafiq Ahmed,1 and Frank G. Rodgers1,{dagger}

National Laboratory for Enteric Pathogens, National Microbiology Laboratory/CSCHAH, Winnipeg, Manitoba,1 Canadian Research Institute for Food Safety, Food Science Department, University of Guelph, Guelph, Ontario, Canada2

Received 18 July 2002/ Returned for modification 29 August 2002/ Accepted 18 October 2002

Salmonella enterica subsp. enterica serotype Enteritidis is not readily subtyped beyond the level of phage type (PT). A recently developed method for ribotyping of this organism, which uses a mixture of PstI and SphI (PS) for restriction of DNA (PS ribotyping), has proved useful for further subtyping of a number of PTs of this organism, including PT 4. However, it has not been extensively tested with PT 8. In the present study the PS ribotyping method was used to investigate outbreaks of both S. enterica serotype Enteritidis PT 4 and PT 8 and provided subtyping data that were consistent with information obtained from epidemiologic investigations. The method proved to be more discriminatory than phage typing and pulsed-field gel electrophoresis (PFGE) combined and was useful for investigating a pseudo-outbreak involving isolates that had identical PTs and PFGE types but that could not be linked epidemiologically. Several PS ribotypes were found within the cluster of isolates indistinguishable by other subtyping methods, confirming the epidemiologic findings. Although the PS ribotyping method proved to have a superior discriminatory ability in resolving clusters, it did not have high enough throughput for use in outbreak investigations. This method has therefore been adapted for use in automated ribotyping with a RiboPrinter, and the results were compared with those obtained by manual ribotyping. Both methods produce equivalent results and are useful for obtaining epidemiologically relevant subtyping data for S. enterica serotype Enteritidis, including PT 8 strains not extensively tested previously.

* Corresponding author. Mailing address: National Laboratory for Enteric Pathogens, National Microbiology Laboratory, Canadian Science Centre for Human and Animal Health, 1015 Arlington St., Winnipeg, Manitoba R3E 3R2, Canada. Phone: (204) 789-2094. Fax: (204) 789-5012. E-mail: Clifford_Clark{at}hc-sc.gc.ca.

{dagger} Present address: Department of Microbiology, University of New Hampshire, Durham, NH 03824-2617.

Journal of Clinical Microbiology, January 2003, p. 27-33, Vol. 41, No. 1
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.1.27-33.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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