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Journal of Clinical Microbiology, February 2007, p. 536-543, Vol. 45, No. 2
0095-1137/07/$08.00+0     doi:10.1128/JCM.01595-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Comparison of Multiple-Locus Variable-Number Tandem Repeat Analysis, Pulsed-Field Gel Electrophoresis, and Phage Typing for Subtype Analysis of Salmonella enterica Serotype Enteritidis{triangledown}

D. Boxrud,1* K. Pederson-Gulrud,1 J. Wotton,1 C. Medus,1 E. Lyszkowicz,2 J. Besser,1 and J. M. Bartkus1

Minnesota Department of Health, Minneapolis, Minnesota,1 Centers for Disease Control and Prevention, Atlanta, Georgia2

Received 2 August 2006/ Returned for modification 28 September 2006/ Accepted 30 November 2006

Strain subtyping is an important tool for detection of outbreaks caused by Salmonella enterica serotype Enteritidis. Current subtyping methods, however, yield less than optimal subtype discrimination. In this study, we describe the development and evaluation of a multiple-locus variable-number tandem repeat analysis (MLVA) method for subtyping Salmonella serotype Enteritidis. The discrimination ability and epidemiological concordance of MLVA were compared with those of pulsed-field gel electrophoresis (PFGE) and phage typing. MLVA provided greater discrimination among non-epidemiologically linked isolates than did PFGE or phage typing. Epidemiologic concordance was evaluated by typing 40 isolates from four food-borne disease outbreaks. MLVA, PFGE, and, to a lesser extent, phage typing exhibited consistent subtypes within an outbreak. MLVA was better able to differentiate isolates between the individual outbreaks than either PFGE or phage typing. The reproducibility of MLVA was evaluated by subtyping sequential isolates from an infected individual and by testing isolates following multiple passages and freeze-thaw cycles. PFGE and MLVA patterns were reproducible for isolates that were frozen and passaged multiple times. However, 2 of 12 sequential isolates obtained from an individual over the course of 36 days had an MLVA type that differed at one locus and one isolate had a different phage type. Overall, MLVA typing of Salmonella serotype Enteritidis had enhanced resolution, good reproducibility, and good epidemiological concordance. These results indicate that MLVA may be a useful tool for detection and investigation of outbreaks caused by Salmonella serotype Enteritidis.

* Corresponding author. Mailing address: Public Health Laboratory, Minnesota Department of Health, 601 Robert Street N., P.O. Box 64899, Saint Paul, MN 55164-0899. Phone: (651) 201-5257. Fax: (601) 201-5070. E-mail: dave.boxrud{at}health.state.mn.us.

{triangledown} Published ahead of print on 6 December 2006.

Journal of Clinical Microbiology, February 2007, p. 536-543, Vol. 45, No. 2
0095-1137/07/$08.00+0     doi:10.1128/JCM.01595-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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