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Journal of Clinical Microbiology, January 2007, p. 81-87, Vol. 45, No. 1
0095-1137/07/$08.00+0     doi:10.1128/JCM.01622-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Genetic Profiling of the Oral Microbiota Associated with Severe Early-Childhood Caries{triangledown}

Y. Li,1* Y. Ge,1,2 D. Saxena,1 and P. W. Caufield1

New York University College of Dentistry, 345 E. 24th Street, New York, New York 10010,1 Peking University School of Stomatology, 22 Zhongguancun Nandajie, Beijing 100081, People's Republic of China2

Received 6 August 2006/ Returned for modification 6 September 2006/ Accepted 13 October 2006

The determination of the composition of the microbial community in the oral cavity is usually based on cultivation methods; however, nearly half of the bacteria in the saliva and the dental plaque are not cultivable. In this study, we evaluated the difference in oral microbial diversity between children with severe early-childhood caries (S-ECC) and caries-free (CF) controls by means of a cultivation-independent approach called denaturing gradient gel electrophoresis (DGGE). Pooled dental plaque samples were collected from 20 children aged 2 to 8 years. Total microbial genomic DNA was isolated from those subjects, and a portion of the 16S rRNA gene locus was PCR amplified by using universal primers. We observed that the mean species richness of the bacterial population was greater in the CF children (n = 12) (42 ± 3.7) than in the S-ECC children (n = 8) (35 ± 4.3); the difference was statistically significant (P = 0.005). The overall diversity of plaque samples as measured by the Shannon index was 3.5 for the S-ECC group and 3.7 for the CF group (P = 0.004). Differences in DGGE profiles were distinguished on the basis of a cluster analysis. Sequence analysis of excised DGGE bands consisted of 2.7 phylotypes, on average. After adjusting for the number of observed bands, we estimated that the S-ECC group exhibited 94.5 total phylotypes and that the CF group exhibited 113.4. These results suggest that the microbial diversity and complexity of the microbial biota in dental plaque are significantly less in S-ECC children than in CF children.

* Corresponding author. Mailing address: Department of Basic Science and Craniofacial Biology, New York University College of Dentistry, 345 E. 24th Street, New York, NY 10010. Phone: (212) 998-9607. Fax: (212) 995-4087. E-mail: yihong.li{at}nyu.edu.

{triangledown} Published ahead of print on 1 November 2006.

Journal of Clinical Microbiology, January 2007, p. 81-87, Vol. 45, No. 1
0095-1137/07/$08.00+0     doi:10.1128/JCM.01622-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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