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Journal of Clinical Microbiology, May 1999, p. 1469-1473, Vol. 37, No. 5
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Detection of Unculturable Bacteria in Periodontal Health and Disease by PCR

R. Harper-Owen,1 D. Dymock,2 V. Booth,3 A. J. Weightman,4 and W. G. Wade1,*

Oral Microbiology Unit1 and Department of Periodontology and Preventive Dentistry,3 King's College London, Guy's Hospital, London SE1 9RT, Division of Oral Medicine, Pathology and Microbiology, Dental School, Bristol BS1 2LY,2 and School of Pure and Applied Biology, University of Cardiff, Cardiff CF1 3TL,4 United Kingdom

Received 28 September 1998/Returned for modification 19 November 1998/Accepted 29 January 1999

Recently developed molecular methods have made it possible to characterize mixed microflora in their entirety, including the substantial numbers of bacteria which do not grow on artificial culture media. In a previous study, molecular analysis of the microflora associated with acute oral infections resulted in the identification of three phylotypes, PUS3.42, PUS9.170, and PUS9.180, representing as-yet-uncultured organisms. The aim of this study was to design and validate specific PCR primers for these phylotypes and to determine their incidences in samples collected from healthy and diseased periodontal tissues. Two specific reverse primers were devised for each phylotype, and these were used in duplex PCRs with universal forward and reverse primers. All three phylotypes were detected in periodontal sites; PUS9.170, related to oral asaccharolytic Eubacterium spp., was significantly associated with disease. This study demonstrates the possibility of using unculturable, and therefore uncharacterized, organisms as markers of disease.

* Corresponding author. Mailing address: Oral Microbiology Unit, Department of Oral Medicine and Pathology, Floor 28, Guy's Tower, Guy's Hospital, London SE1 9RT, United Kingdom. Phone: 171 955 2849. Fax: 171 955 2847. E-mail: w.wade{at}umds.ac.uk.

Journal of Clinical Microbiology, May 1999, p. 1469-1473, Vol. 37, No. 5
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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