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Journal of Clinical Microbiology, December 2003, p. 5389-5397, Vol. 41, No. 12
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.12.5389-5397.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Multilocus Variable-Number Tandem Repeat Analysis Distinguishes Outbreak and Sporadic Escherichia coli O157:H7 Isolates

Anna C. Noller,1,2 M. Catherine McEllistrem,1 Antonio G. F. Pacheco,3,4 David J. Boxrud,5 and Lee H. Harrison1*

Infectious Diseases Epidemiology Research Unit, University of Pittsburgh Graduate School of Public Health and School of Medicine,1 Department of Infectious Diseases and Microbiology,2 Department of Epidemiology, University of Pittsburgh Graduate School of Public HealthSchool of Medicine, Pittsburgh, Pennsylvania,3 Departamento de Epidemiologia e Métodos Quantitativos em Saúde, Escola Nacional de Saúde Pública, FIOCRUZ, Rio de Janeiro, Brazil,4 Microbiology Laboratory, Minnesota Department of Health, Minneapolis, Minnesota5

Received 16 April 2003/ Returned for modification 7 July 2003/ Accepted 11 September 2003

Escherichia coli O157:H7 is a major cause of food-borne illness in the United States. Outbreak detection involves traditional epidemiological methods and routine molecular subtyping by pulsed-field gel electrophoresis (PFGE). PFGE is labor-intensive, and the results are difficult to analyze and not easily transferable between laboratories. Multilocus variable-number tandem repeat (VNTR) analysis (MLVA) is a fast, portable method that analyzes multiple VNTR loci, which are areas of the bacterial genome that evolve quickly. Eighty isolates, including 21 isolates from five epidemiologically well-characterized outbreaks from Pennsylvania and Minnesota, were analyzed by PFGE and MLVA. Strains in PFGE clusters were defined as strains that differed by less than or equal to one band by using XbaI and the confirmatory enzyme SpeI. MLVA was performed by comparing the number of tandem repeats at seven loci. From 6 to 30 alleles were found at the seven loci, resulting in 64 MLVA types among the 80 isolates. MLVA correctly identified the isolates from all five outbreaks if only a single-locus variant was allowed. MLVA differentiated strains with unique PFGE types. Additionally, MLVA discriminated strains within PFGE-defined clusters that were not known to be part of an outbreak. In addition to being a simple and validated method for E. coli O157:H7 outbreak detection, MLVA appears to have a sensitivity equal to that of PFGE and a specificity superior to that of PFGE.

* Corresponding author. Mailing address: Infectious Diseases Epidemiology Research Unit, University of Pittsburgh Graduate School of Public Health and School of Medicine, 521 Parran Hall, 130 DeSoto St., Pittsburgh, PA 15261. Phone: (412) 624-3137. Fax: (412) 624-2256. E-mail: lharriso{at}edc.pitt.edu.

Journal of Clinical Microbiology, December 2003, p. 5389-5397, Vol. 41, No. 12
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.12.5389-5397.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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