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Journal of Clinical Microbiology, April 2002, p. 1181-1187, Vol. 40, No. 4
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.4.1181-1187.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Population Structure and Genetic Diversity of Actinobacillus actinomycetemcomitans Strains Isolated from Localized Juvenile Periodontitis Patients

Jeffrey B. Kaplan, Helen C. Schreiner, David Furgang, and Daniel H. Fine*

Department of Oral Biology, New Jersey Dental School, Newark, New Jersey 07103-2714

Received 23 July 2001/ Returned for modification 3 October 2001/ Accepted 9 November 2001

The phylogeny of 20 Actinobacillus actinomycetemcomitans strains isolated from patients with localized juvenile periodontitis (LJP) was investigated by using partial sequence analysis of 16S rRNA genes, arbitrarily primed PCR (AP-PCR), and four additional PCR assays that amplified polymorphic regions in the leukotoxin (lkt), cytolethal distending toxin (cdt), major fimbrial subunit (flp-1), and serotype-specific O polysaccharide gene clusters. Our analysis also included four strains isolated from healthy subjects and nine reference strains. We found that A. actinomycetemcomitans strains comprised three major phylogenetic lineages. One lineage consisted of serotype b strains, a second lineage consisted of serotype c strains, and a third lineage consisted of serotype a, d, e, and f strains. 16S rRNA sequences within each lineage were highly conserved (<1% base substitutions), whereas sequences between lineages were exceptionally divergent (1.9 to 5.0% substitutions). Two strains exhibited 16S rRNA sequences that were even more distantly related to those of the three major lineages (2.7 to 6.7% substitutions), indicating that additional minor lineages or variants exist. The distribution of 16S rRNA sequences and lkt, cdt, flp-1, and AP-PCR genotypes was consistent with a clonal population structure, with little evidence of assortative recombination between strains of different serotypes. Strains from all three major lineages were recovered from LJP patients, suggesting that phylogenetically diverse strains of A. actinomycetemcomitans carry pathogenic potential.

* Corresponding author. Mailing address: Medical Science Building, Rm. C-636, 185 S. Orange Ave., Newark, NJ 07103-2714. Phone: (973) 972-7053. Fax: (973) 972-0045. E-mail: finedh{at}umdnj.edu.

Journal of Clinical Microbiology, April 2002, p. 1181-1187, Vol. 40, No. 4
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.4.1181-1187.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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