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Journal of Clinical Microbiology, April 2009, p. 1149-1154, Vol. 47, No. 4
0095-1137/09/$08.00+0     doi:10.1128/JCM.01607-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Utility of Multilocus Variable-Number Tandem-Repeat Analysis as a Molecular Tool for Phylogenetic Analysis of Shigella sonnei{triangledown}

Chien-Shun Chiou,1,2* Haruo Watanabe,3 You-Wun Wang,1 Wan-Ling Wang,1 Jun Terajima,3 Kwai-Lin Thong,4 Dac Cam Phung,5 and Sheng Kai Tung1

Central Region Laboratory, Center for Research and Diagnostics, Centers for Disease Control, Taichung City 40855, Taiwan,1 Institute of Medicine, Chung Shan Medical University, Taichung City 40201, Taiwan,2 National Institute of Infectious Diseases, Tokyo, Japan,3 University of Malaya, 50603 Kuala Lumpur, Malaysia,4 Division of Enteric Infections, National Institute of Hygiene and Epidemiology, Vietnam5

Received 19 August 2008/ Returned for modification 5 December 2008/ Accepted 11 February 2009

A panel of 916 isolates, including 703 closely related IST1 isolates, were characterized by inter-IS1 spacer typing (IST), pulsed-field gel electrophoresis (PFGE), and multilocus variable-number tandem-repeat (VNTR) analysis (MLVA) to evaluate the utility of MLVA as a molecular tool for the phylogenetic analysis of Shigella sonnei. The global phylogenetic patterns determined by IST, PFGE, and MLVA were concordant. MLVA was carried out using 26 VNTR loci with a range of degrees of variability. MLVA data for the 703 IST1 isolates revealed that diversification among the closely related isolates was attributed mainly to four highly variable loci. The phylogenetic pattern for the closely related isolates determined using MLVA profiles of 8 highly variable loci was in agreement with that determined using the 26-locus profiles. A clustering analysis using the profiles of 18 loci with limited variability established clear phylogenetic relationships among IST clonal groups. Accordingly, MLVA is a useful tool for the phylogenetic analysis of S. sonnei. Combined VNTR loci with higher variability are useful markers for resolving closely related isolates, whereas combined loci with lower variability are suitable for establishing clear phylogenetic relationships between strains or clones that have evolved over a longer timescale.

* Corresponding author. Mailing address: Central Region Laboratory, Center for Research and Diagnostics, Centers for Disease Control, Taichung City 40855, Taiwan. Phone: 886-4-24750452. Fax: 886-4-24750452. E-mail: nipmcsc{at}cdc.gov.tw

{triangledown} Published ahead of print on 18 February 2009.

Journal of Clinical Microbiology, April 2009, p. 1149-1154, Vol. 47, No. 4
0095-1137/09/$08.00+0     doi:10.1128/JCM.01607-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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