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Journal of Clinical Microbiology, February 2006, p. 374-377, Vol. 44, No. 2
0095-1137/06/$08.00+0     doi:10.1128/JCM.44.2.374-377.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Locus-Specific Mutational Events in a Multilocus Variable-Number Tandem Repeat Analysis of Escherichia coli O157:H7

Anna C. Noller,1,2 M. Catherine McEllistrem,1 Kathleen A. Shutt,1 and Lee H. Harrison1*

Infectious Diseases Epidemiology Research Unit,1 Department of Infectious Diseases and Microbiology, University of Pittsburgh Graduate School of Public Health and School of Medicine, Pittsburgh, Pennsylvania2

Received 2 June 2005/ Returned for modification 17 August 2005/ Accepted 30 November 2005

Multilocus variable-number tandem repeat analysis (MLVA) is a validated molecular subtyping method for detecting and evaluating Escherichia coli O157:H7 outbreaks. In a previous study, five outbreaks with a total of 21 isolates were examined by MLVA. Nearly 20% of the epidemiologically linked strains were single-locus variants (SLV) of their respective predominant outbreak clone. This result prompted an investigation into the mutation rates of the seven MLVA loci (TR1 to TR7). With an outbreak strain that was an SLV at the TR1 locus of the predominant clone, parallel and serial batch culture experiments were performed. In a parallel experiment, none (0/384) of the strains analyzed had mutations at the seven MLVA loci. In contrast, in the two 5-day serial experiments, 4.3% (41/960) of the strains analyzed had a significant variation in at least one of these loci (P < 0.001). The TR2 locus accounted for 85.3% (35/41) of the mutations, with an average mutation rate of 3.5 x 10–3; the mutations rates for TR1 and TR5 were 10-fold lower. Single additions accounted for 77.1% (27/35) of the mutation events in TR2 and all (6/6) of the additions in TR1 and TR5. The remaining four loci had no slippage events detected. The mutation rates were locus specific and may impact the interpretation of MLVA data for epidemiologic investigations.

* 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, February 2006, p. 374-377, Vol. 44, No. 2
0095-1137/06/$08.00+0     doi:10.1128/JCM.44.2.374-377.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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