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Development of Multiple-Locus Variable-Number Tandem-Repeat Analysis for the Molecular Subtyping of Enterobacter sakazakii

Centre for Food Safety,¹ School of Agriculture, Food Science and Veterinary Medicine,² Conway Institute of Biomolecular and Biomedical Research,⁵ School of Public Health and Population Sciences, University College Dublin, Belfield, Dublin 4, Ireland,⁶ Institute for Food Safety and Hygiene, Vetsuisse Faculty, University of Zurich, CH-8057 Zurich, Switzerland,³ Veterinary Food Safety Laboratory and Cork County Council, Inniscara, County Cork, Ireland⁴

Received 26 July 2007/ Accepted 28 November 2007

The genomic content of Enterobacter sakazakii strain ATCC BAA-894 was analyzed for variable-number tandem repeats (VNTRs). In this study we report the development of a multiple-locus VNTR analysis (MLVA) strategy for the subtyping of E. sakazakii. The method is based on a GeneScan analysis of four VNTR loci labeled with multiple fluorescent dyes. This approach was applied to a collection of 112 isolates representing all 16 of the currently defined E. sakazakii biogroups. MLVA successfully discriminated among these isolates and compared favorably with pulsed-field gel electrophoresis. The method was relatively fast and easy to perform. The potential value of MLVA as an epidemiological tool is discussed.

Published ahead of print on 14 December 2007.