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Journal of Clinical Microbiology, December 2004, p. 5523-5527, Vol. 42, No. 12
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.12.5523-5527.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Culture-Independent Identification of Periodontitis-Associated Porphyromonas and Tannerella Populations by Targeted Molecular Analysis

A. de Lillo,1 V. Booth,1 L. Kyriacou,1 A. J. Weightman,2 and W. G. Wade1*

Infection Research Group, Dental Institute, King's College London, London,1 Cardiff School of Biosciences, Cardiff University, Cardiff, United Kingdom2

Received 14 June 2004/ Returned for modification 26 July 2004/ Accepted 13 August 2004

Periodontitis is the commonest bacterial disease of humans and is the major cause of adult tooth loss. About half of the oral microflora is unculturable; and 16S rRNA PCR, cloning, and sequencing techniques have demonstrated the high level of species richness of the oral microflora. In the present study, a PCR primer set specific for the genera Porphyromonas and Tannerella was designed and used to analyze the bacterial populations in subgingival plaque samples from inflamed shallow and deep sites in subjects with periodontitis and shallow sites in age- and sex-matched controls. A total of 308 clones were sequenced and found to belong to one of six Porphyromonas or Tannerella species or phylotypes, one of which, Porphyromonas P3, was novel. Tannerella forsythensis was found in significantly higher proportions in patients than in controls. Porphyromonas catoniae and Tannerella phylotype BU063 appeared to be associated with shallow sites. Targeted culture-independent molecular ecology studies have a valuable role to play in the identification of bacterial targets for further investigations of the pathogenesis of bacterial infections.

* Corresponding author. Mailing address: Department of Microbiology, Floor 28, Guy's Tower, Guy's Hospital, London SE1 9RT, United Kingdom. Phone: 44 20 7188 3872. Fax: 44 20 7188 3871. E-mail: william.wade{at}kcl.ac.uk.

Journal of Clinical Microbiology, December 2004, p. 5523-5527, Vol. 42, No. 12
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.12.5523-5527.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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