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Journal of Clinical Microbiology, November 2008, p. 3626-3635, Vol. 46, No. 11
0095-1137/08/$08.00+0     doi:10.1128/JCM.00300-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Comparison of Four Molecular Typing Methods for Characterization of Corynebacterium diphtheriae and Determination of Transcontinental Spread of C. diphtheriae Based on BstEII rRNA Gene Profiles{triangledown}

Aruni De Zoysa,1* Peter Hawkey,2 Andre Charlett,3 and Androulla Efstratiou1

Respiratory and Systemic Infection Laboratory, Health Protection Agency, Centre for Infections, 61 Colindale Avenue, London NW9 5EQ, United Kingdom,1 Health Protection Agency, Heartlands Hospital, Birmingham, United Kingdom, and Division of Immunity and Infection, Medical School, University of Birmingham, Birmingham, United Kingdom,2 Statistics, Modelling & Bioinformatics, Health Protection Agency, Centre for Infections, 61 Colindale Avenue, London NW9 5EQ, United Kingdom3

Received 13 February 2008/ Returned for modification 8 May 2008/ Accepted 2 September 2008

The diphtheria epidemic in the Russian Federation in the 1990s made diphtheria a focus of global concern once again. The development of rapid and reproducible typing methods for the molecular characterization of Corynebacterium diphtheriae has become a priority in order to be able to monitor the spread of this important pathogen on a global scale. We report on a comparison of four molecular typing methods (ribotyping, pulsed-field gel electrophoresis [PFGE], random amplification of polymorphic DNA [RAPD], and amplified fragment length polymorphism [AFLP]) for the characterization of C. diphtheriae strains. Initially, 755 isolates originating from 26 countries were analyzed by ribotyping. One strain of each ribotype was then randomly chosen and characterized by PFGE, RAPD, and AFLP. In order to ascertain whether the Eastern European epidemic ribotype could be further discriminated, 10 strains of ribotype D1 (the epidemic ribotype) from different geographical regions were randomly chosen and subjected to analysis by PFGE, RAPD, and AFLP. The results revealed that ribotyping is highly discriminatory and reproducible and is currently the method of choice for typing C. diphtheriae. PFGE and AFLP were less discriminatory than ribotyping and RAPD. An assessment of the transcontinental spread of the organism showed that several genotypes of C. diphtheriae circulated on different continents of the world and that each outbreak was caused by a distinct clone. The ribotypes seen in Europe appeared to be distinct from those seen elsewhere, and certain ribotypes appeared to be unique to particular countries.

* Corresponding author. Mailing address: Systemic Infection Laboratory, Health Protection Agency Centre for Infections, 61 Colindale Avenue, London NW9 5EQ, United Kingdom. Phone: 44 208-327-7536. Fax: 44 208-200-6528. E-mail: aruni.dezoysa{at}hpa.org.uk

{triangledown} Published ahead of print on 10 September 2008.

Journal of Clinical Microbiology, November 2008, p. 3626-3635, Vol. 46, No. 11
0095-1137/08/$08.00+0     doi:10.1128/JCM.00300-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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