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Journal of Clinical Microbiology, February 2005, p. 843-849, Vol. 43, No. 2
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.2.843-849.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Molecular Analysis of Microbial Diversity in Advanced Caries

Kim-Ly Chhour,1 Mangala A. Nadkarni,1 Roy Byun,1 F. Elizabeth Martin,1,2 Nicholas A. Jacques,1 and Neil Hunter1*

Institute of Dental Research, Westmead Millennium Institute and Westmead Centre for Oral Health, Westmead,1 Faculty of Dentistry, The University of Sydney, Sydney, New South Wales, Australia2

Received 13 May 2004/ Returned for modification 15 July 2004/ Accepted 24 September 2004

Real-time PCR analysis of the total bacterial load in advanced carious lesions has shown that the total load exceeds the number of cultivable bacteria. This suggests that an unresolved complexity exists in bacteria associated with advanced caries. In this report, the profile of the microflora of carious dentine was explored by using DNA extracted from 10 lesions selected on the basis of comparable total microbial load and on the relative abundance of Prevotella spp. Using universal primers for the 16S rRNA gene, PCR amplicons were cloned, and approximately 100 transformants were processed for each lesion. Phylogenetic analysis of 942 edited sequences demonstrated the presence of 75 species or phylotypes in the 10 carious lesions. Up to 31 taxa were represented in each sample. A diverse array of lactobacilli were found to comprise 50% of the species, with prevotellae also abundant, comprising 15% of the species. Other taxa present in a number of lesions or occurring with high abundance included Selenomonas spp., Dialister spp., Fusobacterium nucleatum, Eubacterium spp., members of the Lachnospiraceae family, Olsenella spp., Bifidobacterium spp., Propionibacterium sp., and Pseudoramibacter alactolyticus. The mechanisms by which such diverse patterns of bacteria extend carious lesions, including the aspect of infection of the vital dental pulp, remain unclear.

* Corresponding author. Mailing address: Mailing address: Institute of Dental Research, Westmead Centre for Oral Health, PO Box 533, Wentworthville, NSW 2145, Australia. Phone: 612-9845-8762. Fax: 612-9845-7599. E-mail: nhunter{at}dental.wsahs.nsw.gov.au.

Journal of Clinical Microbiology, February 2005, p. 843-849, Vol. 43, No. 2
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.2.843-849.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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