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Journal of Clinical Microbiology, June 2002, p. 1994-2001, Vol. 40, No. 6
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.6.1994-2001.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Multilocus Sequence Typing of Bordetella pertussis Based on Surface Protein Genes

Inge H. M. van Loo,1,2 Kees J. Heuvelman,1 Audrey J. King,1 and Frits R. Mooi1,2*

Laboratory for Infectious Diseases Research, National Institute of Public Health and the Environment, Bilthoven,1 Eijkman Winkler institute for Medical Microbiology, University Medical Center Utrecht, Utrecht, The Netherlands2

Received 29 October 2001/ Returned for modification 1 January 2002/ Accepted 19 March 2002

Despite more than 50 years of vaccination, Bordetella pertussis has remained endemic in The Netherlands, causing epidemic outbreaks every 3 to 5 years. Strain variation may play a role in the persistence of B. pertussis and was studied by sequencing 15 genes coding for surface proteins, including genes for all five components of acellular pertussis vaccines: pertussis toxin (Ptx), pertactin (Prn), filamentous hemagglutinin, and fimbriae (Fim2 and Fim3). A low level of allelic variation was observed, confirming a recent evolutionary origin of B. pertussis. In modern isolates, polymorphism was observed only in prn, ptxS1, ptxS3, and tcfA. Polymorphism in ptxS1, ptxS3, and tcfA was used to categorize isolates in multilocus sequence types (MLSTs). Analysis of Dutch isolates from 1949 to 1999 revealed five MLSTs, which showed a highly dynamic temporal behavior. We observed significant changes in the MLSTs after the introduction of pertussis vaccination in The Netherlands. Epidemic years were found to be associated with the expansion of MLST-4 or MLST-5. MLST-5 showed a remarkable expansion from 10% in 1997 to 80% in 1999. The MLST analysis was extended to a number of widely separated geographic regions: Finland, Italy, Japan, and the United States. MLST-4 and MLST-5 were found to dominate in Italy and the United States. In Finland and Japan, MLST-3 and MLST-2, respectively, were predominant. Thus, although each region showed distinctive MLST frequencies, in three of the five regions MLST-4 and MLST-5 were predominant. These types may represent newly emerged, successful clones. The identification of highly successful clones may shed light on the question of how B. pertussis is able to maintain itself in vaccinated populations.

* Corresponding author. Mailing address: Laboratory for Infectious Diseases Research, National Institute of Public Health and the Environment, P.O. Box 1, 3720 BA Bilthoven, The Netherlands. Phone: 31-30-2743091. Fax: 31-30-2744449. E-mail: fr.mooi{at}rivm.nl.

Journal of Clinical Microbiology, June 2002, p. 1994-2001, Vol. 40, No. 6
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.6.1994-2001.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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